TW201211533A - Microfluidic device for simultaneous detection of multiple conditions in a patient - Google Patents

Abstract

A microfluidic device for simultaneous detection of multiple conditions in a patient, the microfluidic device having an inlet for receiving a sample of biological material drawn from the patient, a microsystems technologies (MST) layer with a detection section having an array of probes for reaction with target molecules in the sample to form probe-target complexes, the target molecules being indicative of medical conditions in the patient, and, a photosensor for detecting the probe-target complexes, wherein, the array of probes has more than 1000 probes.

Description

201211533 VI. Description of the Invention: TECHNICAL FIELD OF THE INVENTION The present invention relates to a diagnostic apparatus using microsystem technology (MST). In particular, the present invention relates to the processing and analysis of microfluidics and biochemicals for molecular diagnostics. [Prior Art] φ molecular diagnostics have been used to provide an early indication of disease detection before the onset of symptoms. Molecular diagnostic tests are used to detect: • genetic disorders • acquired diseases • infectious diseases • genes associated with health-related genetic factors due to high accuracy and rapid processing time, molecular diagnostic tests can reduce the incidence of ineffective health care, Improve patient outcome, improve disease management, and individualized patient care. Many techniques for molecular diagnostics are based on the detection and identification of specific nucleic acids (both deoxyribonucleic acid (DNA) and ribonucleic acid (RNA)) extracted and amplified from biological samples such as blood or saliva. The complementary nature of the nucleobase allows the short sequence (oligonucleotide) of the synthesized DNA to bind (hybridize) to a particular nucleic acid sequence for use in nucleic acid assays. If hybridization occurs, the complementary sequence is present in the sample. This makes it possible, for example, to predict the disease that an individual will get in the future, to determine the type and pathogenicity of an infectious pathogen, or to determine an individual's response to a drug. -5- 201211533 Nucleic Acid-Based Molecular Diagnostic Tests Nucleic acid-based assays have four separate steps: 1. Sample preparation 2. Nucleic acid extraction 3. Nucleic acid amplification (optional) 4. Detection of many sample types, such as blood , urine, sputum and tissue samples for genetic analysis. Diagnostic tests determine the type of sample required, as not all samples represent disease progression. These samples have various components, but usually only one of them is of interest. For example, in the blood, high concentrations of red blood cells can inhibit the detection of pathogenic microorganisms. Therefore, purification and/or concentration steps are typically required at the beginning of the nucleic acid assay. Blood is one of the most frequently requested sample types. It has three main components: white blood cells, red blood cells, and platelets. Platelets accelerate coagulation and maintain activity outside the body. To inhibit coagulation, the sample was mixed with a reagent such as ethylenediaminetetraacetic acid (EDTA) prior to purification and concentration. Red blood cells are typically removed from the sample to concentrate the target cells. In the human body, red blood cells account for about 99% of cellular material, but they do not carry DNA because they do not have a nucleus. In addition, red blood cells contain components such as heme that may interfere with downstream nucleic acid amplification procedures (described below). Red blood cell removal can be accomplished by dissolving the solution differentially red blood cells, leaving the entire remaining cellular material, which can then be separated from the sample by centrifugation. This provides a concentrated target cell from which the nucleic acid is extracted. The exact procedure used to extract nucleic acids depends on the sample and the diagnosis to be performed -6- 201211533 Analysis. For example, the protocol used to extract viral RNA is quite different from the protocol used to extract genomic DNA. However, extracting nucleic acids from a target cell typically involves a cell lysis step and subsequent nucleic acid purification. The cell lysis step ruptures the cell and nuclear membrane and releases the genetic material. This is often done using a lysing detergent such as sodium sulfoxide, which also denatures a large amount of protein present in the cells. The nucleic acid is then purified by an alcohol precipitation step, which is usually carried out using glacial alcohol or isopropanol, or by a solid phase purification step, usually in a column of cerium oxide substrate, resin or It is carried out on paramagnetic beads in the presence of a high concentration of chaotropic salts, followed by washing of the nucleic acids, followed by elution with a buffer of low ionic strength. A selective step prior to precipitation of the nucleic acid is the addition of a protease which digests the protein to further purify the sample. Other lysis methods include mechanical lysis via ultrasonic vibration and heating of the sample to 94t to disrupt thermal lysis of the cell membrane. The φ target DNA_ or RNA can be present in the extracted material in very small amounts, especially if the target is from a pathogenic source. Nucleic acid amplification provides the ability to selectively amplify (i.e., replicate) a particular target in a low concentration to a detectable amount. 〇 The most commonly used nucleic acid amplification technique is the polymerase chain reaction (PCR). PCR is well known in the art, and a complete description of this type of reaction is provided in E. van Pelt-Verkuil et al. Principles and Technical Aspects of PCR Amplification, Springer, 2008 〇 PCR is a useful technique. The 201211533 target DNA sequence was amplified in the context of complex DNA. If you want to amplify rna (by PCR), you must first use an enzyme called reverse transcriptase to transcribe RNA into cDNA (complementary DNA). Subsequently, the obtained cdnA was amplified by PCR. PCR is an exponential method and can be continued as long as the conditions for maintaining the reaction are acceptable. The components of the PCR reaction are:

1. A short single strand of DN A 2. DNA polymerase-synthesized DNA thermostable enzyme having a nucleotide complementary to a region of about 10 to 30 flanking target sequences 3. Deoxyribose Nucleoside triphosphate (dNTP) - provides nucleotides that are incorporated into newly synthesized DNA strands. 4. Buffer - provides the ideal chemical environment for DNA synthesis. PCR typically involves placing these reactants in a vial containing about the extracted nucleic acid (about 1 to 50 microliters). The tube is placed in a thermal cycler; the reactor is a device that allows the reaction to take unequal time at a range of different temperatures. The standard procedures for each thermal cycle involve the denaturing phase, the annealing phase, and the extended phase. The extension phase is sometimes referred to as the primer extension phase. In addition to these three-step procedures, a two-step thermal procedure can also be used in which the annealing phase and the extension phase are combined. The denatured phase typically involves raising the reaction temperature to 90 to 95 °C to denature the DN A strand; in the annealed phase, the temperature is reduced to 50 to 6 (TC to bond the primer; then in the extended phase, the temperature is raised The optimal DNA polymerase activity temperature is 60 to 72 ° C for primer extension. This process is repeated about 20 to 40 times, and the final result is to generate millions of copies of the target sequence between the primers. Variants of many standard PCR protocols for molecular diagnostics, -8- 201211533 These include, for example, multiple primer set PCR, linker-primed PCR, direct PCR, tandem PCR, real-time PCR, and reverse transcription Enzyme PCR Multi-initiator PCR uses multiple primer sets in a single PCR mix to generate different sizes of amplicons specific for different DNA sequences. By single target (targeting) multiple genes, one can experiment from a single experiment. Get extra information (in other ways, it takes several trials). The most φ optimization of multi-initiator PCR is more difficult because it requires the selection of primers with approximate adhesion temperature and the extension of approximate length and base composition. To ensure equal amplification efficiency of each amplicon. Linker-primed PCR, also known as ligation adaptor PCR, is a type that does not require a target-specific primer. A method of amplifying a nucleic acid of substantially all DNA sequences in a complex DN A mixture. This method first digests the target DNA population with a suitable restriction endonuclease (enzyme). The ligase will then have a suitable suspension. The φ double-stranded oligonucleotide linker (also known as a conjugate) is ligated to the end of the target DNA fragment. Next, nucleic acid amplification is performed using an oligonucleotide primer having specificity for the linker sequence. Thus, all DNA-derived fragments contiguous with the linker oligonucleotide can be amplified. Direct PCR describes a system that performs PCR directly on a sample without any (or minimal) nucleic acid extraction. Many components present in the purified biological sample, such as the protohemoglobin component in the blood, inhibit the PCR reaction. Therefore, it is customary to enhance the purification of the target nucleic acid before preparing the PCR reaction mixture. Chemical properties and appropriate changes in sample concentration -9-201211533 degrees, it is possible to perform minimal DNA purification or PCR by direct PCR. The PCR chemistry for direct PCR involves increasing buffer strength and using high activity. Pr〇cessivity polymerase and addition to potential polymerase inhibitors. Repeat PCR utilizes two separate nucleic acid amplification cycles to increase the probability of amplifying the correct amplicon. One type is nested PCR, in which two pairs of PCR primers are used to amplify a single locus in different nucleic acid amplification cycles. The first pair of primers hybridize to a nucleic acid sequence located in a region other than the target nucleic acid sequence. A second pair of primers (nested primers) is used for the second amplification, the pair of primers being incorporated within the first PCR product to produce a second PCR product comprising the target nucleic acid, and the second product is short. The rationale used in this strategy is that if the wrong locus is amplified due to a mistake during the first nucleic acid amplification, the probability that the wrong locus is re-amplified by the second pair of primers is very low, thus ensuring specificity. Real-time PCR or quantitative PCR is used to measure the amount of PCR product in real time. The initial amount of nucleic acid in the sample can be determined by using a probe containing a fluorophore or a fluorescent dye and a set of standards in the reaction. This is especially useful for molecular diagnostics where the choice of treatment may vary depending on the amount of pathogen in the sample. Reverse transcriptase PCR (RT-PCR) is used to amplify DNA from RNA. The reverse transcriptase is an enzyme that reverse transcribes RNA into complementary DNA (cDNA), which is then amplified by PCR. RT-PCR is widely used for expression profiling to determine the expression of a gene or to identify sequences of an RNA transcript, including transcription initiation and termination sites. It is also used to amplify -10- 8 201211533 RNA viruses, such as human immunodeficiency virus or hepatitis C virus. The isothermal amplification is another form of nucleic acid amplification that does not rely on the thermal denaturation of the target DNA during the amplification reaction, thus eliminating the need for sophisticated instruments. Therefore, the thermostatic nucleic acid amplification method can be carried out in a field place or simply in an environment other than the laboratory. Some thermostatic nucleic acid amplification methods have been described, including Strand Displacement Amplification, Transcription Mediated Amplification, Nucleic Acid Sequence Based Amplification, Recombinase Polymerase Expansion Recombinase Polymerase Amplification, Rolling Circle Amplification, Ramification Amplification, Helicase-Dependent Thermostatic DNA Amplification (H e 1 icase - D ep end en t Isothermal DNA Amplification Loop-Mediated Isothermal Amplification o Constant-temperature nucleic acid amplification does not rely on continuous heating to denature template DNA to produce φ as a template for continued amplification of the template, but to generate a single using other methods at constant temperature. A strand of a molecule, such as enzymatic cleavage of a DNA molecule by specific restriction of an endonuclease, or separation of a DNA double strand by an enzyme. Strand-substituted amplification (SDA) relies on the ability of specific restriction enzymes to cleave unmodified strands of hemi-m〇dified DNA, and polymerase extensions that lack 5'-3' exonuclease activity and replace downstream stocks Ability. Exponential nucleic acid amplification is then achieved by coupling sense and antisense, wherein the strand from the sense reaction replaces the template for the antisense reaction. The nicking enzyme used in this reaction does not cleave the DNA in a conventional manner, but instead produces an incision in one of the -11 - 201211533 DNA strands, such as N.Alwl, N.BstNBl, and Mlyl. SDA is improved by using a combination of a thermostable restriction enzyme (Aval) and a thermostable external polymerase (B st polymerase). This combination has been shown to increase the amplification efficiency of the reaction from 1-8 fold amplification to 101Q fold amplification, so it is possible to utilize this technique to amplify unique single copy molecules. Transcription-mediated amplification (TMA) and nucleic acid sequence-based amplification (NASB A) RNA polymerase is used to replicate RNA sequences rather than corresponding genomic DNA. This technique uses two primers and two or three enzymes, RNA polymerase, reverse transcriptase, and selective RNase Η (if reverse transcriptase does not have Rnase activity). One of the primers contains a promoter sequence of RNA polymerase. In the first step of nucleic acid amplification, the primer hybridizes to the target ribosomal RNA (rRNA) at a defined site. The reverse transcriptase then extends from the 3' end of the promoter primer to produce a DNA copy of the target rRNA. The RNA in the formed RNA:DNA dimer is decomposed by the RNase activity (if any) of the reverse transcriptase or additional RN a se. In the next step, the second primer binds to the DNA copy. The new DNA strand is synthesized by reverse transcriptase from the end of this primer to produce a double-stranded DNA molecule. RNA polymerase recognizes the promoter sequence in the DNA template and initiates transcription. Each newly synthesized RNA expander is re-entered as a template for a new replication cycle. In recombinase polymerase amplification (RPA), isothermal amplification of a specific DNA fragment is achieved by binding an oppositely directed oligonucleotide primer to the template DNA and then extending the primers by a DNA polymerase. . Denaturation of the double-stranded D N A (d s D N A) template does not require heating. Instead, RPA uses a recombinase-introduction complex to scan dsDNA to facilitate stock exchange at the homologous 201211533 locus. The resulting structure is stabilized by the interaction of a single strand of DNA binding protein with the substituted template strand, thereby preventing the primer from exiting via branch migration. Recombinase unwinding allows the strand-substituted DNA polymerase (such as a large fragment of B. subtilis P〇l I (BSU)) to be accessible to the 3' end of the oligonucleotide, followed by primer extension. Exponential nucleic acid amplification is accomplished by repeated cycles of this process. The helicase-dependent amplification (HDA) mimics an in vivo system in which φ is used with DNA helicase to generate a single-strand template for primer hybridization, followed by extension of the primer by DNA polymerase. In the first step of the HDA reaction, the de-enzyme is passed along the target DNA to interrupt the hydrogen bond between the two strands, which in turn binds to the single-stranded binding protein. The primers are bonded by exposing the single-strand target region by helicase. The DNA polymerase then uses the free deoxyribonucleoside triphosphate (dNTP) to extend the 3' end of each primer to make two DNA replicas. The two dsDNA replication strands each enter the next HDA cycle, resulting in amplification of the exponential nucleic acid of the target sequence. φ Other DNA-based thermostating techniques include rolling circle amplification (RCA) in which a DNA polymerase extends the primer around a circular DNA template to produce a long DNA product consisting of many repeat copies of the loop. Prior to the end of the reaction, the polymerase produced tens of thousands of copies of the circular template and the copies of the strands were tethered to the original target DNA. This approach allows for spatial dissociation of the target and rapid nucleic acid amplification of the signal. A copy of 1 to 12 templates can be generated in one hour. A variant of the branched-type amplification system RCA that uses a closed circular probe (C-probe) or a padlock probe and a DNA polymerase with high processivity to exponentially expand under constant temperature conditions - 13- 201211533 The C-probe. Circular thermostat amplification (LAMP) provides high selectivity using dnA polymerase and a set of four specially designed primers that recognize a total of different sequences on the standard DNA. The primers within the sense and antisense strand sequences containing the standard DNA initiate LAMP. The strands initiated by the exogenous primers then replace the DNA synthesis to release a single strand of DNA. The single-stranded DNA can serve as a template for DNA synthesis initiated by a second primer and an external primer, and the second primer and the foreign primer are hybridized to the other end of the standard IE, and the DNA is synthesized to produce stem-loop DNA. structure. In the subsequent LAMP cycle, an internal primer hybridizes to the loop on the product and initiates a substitutional DN A synthesis, producing the original stem-loop DNA and a new rate of loop DNA with twice as long stems. The cycle reaction continued to accumulate 1 09 copies of the target within - hours. The final product is a stem-loop DNA having a plurality of inverted repeats of the target and a cauliflower-like structure, wherein the plurality of loops in the cauliflower-like structure are formed by adhering mutually overlapping target repeats in the same strand. . After completion of nucleic acid amplification, the amplification product must be analyzed to determine whether or not the expected amplicon (amplification amount of the target nucleic acid) is produced. The method of analyzing the product can be carried out by simply measuring the size of the amplicon by gel electrophoresis to hybridization with DN A to identify the nucleotide composition of the amplicon. Gel electrophoresis is the easiest way to check whether a nucleic acid amplification method produces the desired amplicon. Gel electrophoresis utilizes an electric field applied to a gel matrix to separate DNA fragments. Negatively charged DNA fragments will move in the matrix at different rates, depending on the fragment size. After the electrophoresis is completed, the fragments in the gel are stained to visualize them. The commonly used dyeing agent for ethidium bromide is -14- 8 201211533. It shows fluorescence under ultraviolet light. The size of the fragments is determined by comparison with DNA ladders containing DNA fragments of known size and migrating side by side with the amplicons on the gel. Since the oligonucleotide primer binds to a specific site adjacent to the target DNA, the size of the amplification product can be predicted and detected as a band of known size on the gel. In order to determine the correctness of the amplicon, or if several amplicons are produced, a DNA probe that hybridizes to the amplicon is typically employed. DNA hybridization refers to the formation of double stranded DNA by complementary base pairing. DNA hybridization of about 20 nucleotides in length is required for DNA hybridization to clearly identify a particular amplification product. If the probe has a sequence complementary to the amplicon (target) DNA sequence, hybridization will occur at the appropriate temperature, pH and ion concentration. If hybridization occurs, the gene or DNA sequence of interest is present in the original sample. Optical detection systems are most commonly used to detect hybridization methods. One of the amplicon and probe φ is labeled with a fluorescent agent or an electrochemical luminescent agent to emit light. These methods differ in that they cause the photo-generating group to produce an excited state, but both can be used to covalently label nucleotide strands. In the case of electrochemiluminescence (ECL), the light system is generated by current stimulating luminescent group molecules or complexes. In the case of fluorescence, it is irradiated with excitation light to cause light to be emitted. The fluorescent system is detected by a light source and a detection unit that provides excitation light of a wavelength absorbed by the fluorescent molecule. The detection unit includes a light sensor (such as a photomultiplier tube or a charge coupled device (C CD) array) that detects the transmitted signal and prevents the excitation light from being included in the output of the light sensor -15-201211533 (such as wavelength Select filter). The fluorescent molecules emit Stokes shifted light in response to the excitation light, which is collected by the detection unit. The Stokes shift is the difference in frequency or wavelength between the emitted light and the absorbed excitation light. The ECL emission system is detected using a light sensor that is sensitive to the emission wavelength of the ECL species used. For example, transition metal-ligand complexes emit light of a visible wavelength, so conventional photodiodes and CCDs can be used as light sensors. An advantage of ECL is that if the ambient light is shielded, the ECL emits light that is the only light in the detection system, thus increasing sensitivity. Microarrays allow hundreds of thousands of DNA hybridization experiments to be performed simultaneously. Microarrays are powerful molecular diagnostic tools that screen thousands of genetic diseases or detect the presence of numerous infectious pathogens in a single experiment. The microarray consists of a number of different DNA probes that are fixed at the point of acceptance. The target DNA (amplicon) is first labeled with fluorescent or luminescent molecules (either during nucleic acid amplification or after nucleic acid amplification), followed by application of the target DNA to the probe microarray. The microarray is cultured in a temperature controlled, humidified environment for hours or days to cause hybridization between the probe and the amplicon. After incubation, the microarray must be washed through a series of buffers to remove unbound strands. After cleaning, the microarray surface is dried with a gas stream (usually nitrogen). The rigor of hybridization and cleaning is critical. Insufficient stringency may result in highly non-specific binding. Excessive rigor may result in inability to properly combine, resulting in reduced sensitivity. Hybridization is identified by detecting the light emitted by the labeled amplicon of the hybridizing probe with the complementary probe. -16- 201211533 Fluorescence from microarrays is detected by a microarray scanner, which is usually a computer-controlled inverted scanning fluorescent conjugated focus microscope that typically uses laser-excited fluorescent dyes and light-sensing The transmitter (such as a photomultiplier tube or CCD) detects the transmitted signal. The fluorophore emits a Stokes bit shift (as described above) which is collected by the detection unit. The emitted fluorescent light must be collected, separated from the unabsorbed excitation wavelength and transmitted to the detector. In a microarray scanner, a conjugate focal configuration of a conjugate focal hole aperture mounted on the φ image plane is typically used to eliminate out-〇f-focus information. This device allows only the light of the focused portion to be detected. Light from above and below the focal plane of the target cannot enter the detector, thus increasing the signal to noise ratio. The detected fluorescent photons are converted into electrical energy by a detector and then converted into a digital signal. This digital signal is translated into a number that represents the intensity of the fluorescence from a given pixel. Each feature of the array consists of one or more of these pixels. The final result of the scan is an image of the surface of the array. Since the exact sequence and position of each probe on the microarray is known, the target sequence to which it is hybridized can be simultaneously identified and analyzed. More information on fluorescent probes can be found at: http://www.premierbiosoft.com/tech_notes/FRET_probe.html and http://www.invitrogen.com/site/us/en/home/References/Molecular- Probes-The-

Handbook/Technical-Notes-and-Product-Highlights/Fluorescence-Resonance-

Energy-Transfer-FRET.html Fixed-point care 0〇1>^-(^-〇八1^) Molecular diagnosis Although molecular diagnostic tests offer many benefits, the growth of such tests in the clinical laboratory of Lin-17-201211533 is still relatively high. The slowness of expectations is not the mainstream of laboratory medical testing. This is primarily due to the higher complexity and cost of nucleic acid detection compared to assays that do not involve nucleic acid methods. The extensive use of molecular diagnostic testing in clinical settings is closely related to the development of instrumentation, which must significantly reduce costs, provide rapid (automatic) analysis from initial (sample processing) to final (resulting), and does not require The operation of substantial human intervention. Point-of-care technology can provide care in the physician's office, on the hospital bed side, or even in a consumer-focused home environment. This technology offers many advantages including: • Quick results, immediate treatment and improved care quality • Very self-contained A small number of samples to obtain the number of laboratories • Reduce clinical workload • Reduce laboratory workload and reduce administrative efficiency to improve office efficiency • Reduce hospitalization days, outpatients can be diagnosed at the initial diagnosis and reduced sample processing , storage and delivery to improve the cost per patient • Helps clinical management decisions such as infection control and antibiotic use. Laboratory wafer (LOC)-based molecular diagnostics provide micro-fluid technology-based molecular diagnostic systems that can be automated and A device that accelerates molecular diagnostic analysis. The shorter detection time is primarily due to the fact that the required sample volume is less active, automated, and low-cost built-in cascaded diagnostic method steps within the microfluidic device. The volume of nanoliters and microliters is also 8 -18- 201211533 to reduce reagent consumption and cost. Laboratory wafer (LOC) devices are a common form of microfluidic device. The LOC device has an MST structure within the MST layer for integrating fluid processing onto a single supporting substrate (usually helium). The unit cost of each LOC device is very low by manufacturing the VLSI (very large integrated circuit) lithography technology of the semiconductor industry. However, controlling the flow of fluid through the LOC device, adding reagents, controlling reaction conditions, and the like requires a large external hydroelectric engineering device. Connecting LOC devices to these external devices is significantly limited to φ1〇 (: molecular diagnostics of the device is used in a laboratory environment. The cost of external instruments and their operational complexity excludes LOC-based molecular diagnostics as a point-of-care environment. In view of this, there is a need for a molecular diagnostic system based on a LOC device for use in point-of-care. [Invention] Various aspects of the present invention are now described by the following numbered paragraphs. φ GAS080.1 This aspect of the invention Provided is a wafer on-lab (LOC) device for detecting a target nucleic acid sequence in a sample, the LOC device comprising: a nucleic acid sequence having a hybrid complementary to the target nucleic acid sequence for use in forming a probe-target hybrid and electrochemiluminescence (ECL) a probe of a luminophore; and an electrode for generating an excited state of the ECL luminophore, wherein the ECL luminophore emits photons of light in the excited state. GAS 08 0.2 Preferably, each probe has Resonance energy transfer to quench a functional portion of photon emission from the ECL luminophore. -19- 201211533 GAS 080.3 Preferably, the probe is configured to When the probe forms a probe-target hybrid, the functional moiety for quenching photon emission from the ECL luminophore is further derived from the ECL luminophore. GAS 080.4 Preferably, the LOC device also has a A CMOS circuit configured to provide electrical pulses to the electrodes. GAS 080.5 Preferably, the electrical pulse has a period of less than 0.69 seconds. GAS 080.6 preferably 'the current of the electrical pulse is between 0.1 Nai and 69.0 Preferably, the electrodes have an anode and a cathode each having a finger-like configuration, the finger structures being configured to intersect the fingers of the anode with the fingers of the cathode. GAS 080.8 Preferably The anode and the cathode are separated by a dielectric gap of 0.4 micrometers to 2.0 micrometers wide. GAS080.9 preferably 'the luminophore organometallic complex. GAS080.1 0 Preferably, the organometallic is misaligned Preferably, the CMOS circuit has a light sensor for sensing photons emitted by the ECL luminophore. GAS 080.1 2 Preferably, the LOC device Also having an array of hybrid chambers, each of which The hybridization chambers each have a pair of electrodes and contain a plurality of probes, and the nucleic acid sequences of the probes in each of the hybridization chambers are different from the nucleic acid sequences in at least one other hybridization chamber in the array, such that the plurality of labeled nucleic acid sequences are Detected. -20- 201211533 GAS080.1 3 Preferably, the LOC device also has a support substrate, wherein the CMOS circuit is located between the hybridization chamber and the support substrate to make the photosensor adjacent to the hybrid Preferably, the light sensor is an array of photodiodes, the position of the photodiode array being configured such that each of the photodiodes respectively corresponds to one of the hybrid chambers By. GAS080.1 5 Preferably, the photodiodes have planar active surface regions for receiving φ light from the luminophore, each of the active surface regions being in the same plane, and the electrodes are patterned by one layer Patterned to form the conductive material of the separate anode and cathode, the plane of the layer being parallel to the plane of the active surface region of the photodiodes 〇GAS080.1 6 preferably, each of the electrodes is centered One of the electrodes causes oxidation or reduction of the luminophore to produce a working electrode of an excited species that emits photons, the working electrode being positioned such that the probes are between the photodiode and the working electrode between. GAS080.1 7 Preferably, the photodiodes have a planar active surface region for receiving light from the luminophore, and the working electrode has a surface region optically coupled to an active surface region of the photodiode The working electrode is configured such that a surface area optically coupled to the active surface area is greater than 50% of an active surface area of the photodiode. GAS 080.1 8 Preferably, the LOC device also has a polymerase interlocking reaction (PCR) portion for amplifying the target nucleic acid sequence in the sample. -21 - 201211533 GAS080.1 9 Preferably, the PCR portion has a heater element for heating the nucleic acid sequence circulating the target and a polymerase, the heater element being configured to be operationally controlled by the CMOS circuit. GAS080.20 Preferably, the LOC device also has a plurality of sensors coupled to the CMOS circuit for feedback control of the electrodes and the heater elements. Electrochemiluminescence has the advantage of producing effective light at controlled locations in a microfluidic environment. In addition, synchronizing with the sensor is more convenient than a technology such as fluorescent. This LOC device has the benefit of less complex design and manufacturing requirements and will therefore result in a simpler, more reliable manufacturing that will detect the target DN 更 more sensitively and more specifically. GAS08 1 · 1 This aspect of the invention provides a wafer-on-lab (LOC) device for detecting a target nucleic acid sequence in a sample, the LOC device comprising: a support substrate; having complementary to the target nucleic acid sequence for formation a probe-target hybrid nucleic acid sequence and a probe for an electrochemiluminescence (ECL) organometallic complex; and an electrode for generating an excited state of the organometallic complex in which the organometallic is misaligned a photon that emits light; and a CMOS circuit between the support substrate and the probes for applying a voltage to the electrodes. GAS08 1.2 Preferably, each of the probes has a functional moiety for quenching photon emission from the organometallic complex by resonance energy transfer. 8-22-201211533 GAS08 1.3 Preferably, the probe is configured The functional moiety for quenching photon emission from the organometallic complex is further derived from the organometallic complex when the probe forms a probe-target hybrid. GAS 081.4 Preferably, the CMOS circuit is configured to provide electrical pulses to the electrodes. φ GAS081.5 Preferably, the electrical pulse has a period of less than 0.69 seconds. Preferably, the electrical current of the electrical pulse is between 0.1 nanoamperes and 69.0 nanoamperes. GAS 081.7 Preferably, the electrodes have an anode and a cathode, each having a finger configuration, the finger structures being configured to intersect the fingers of the anode with the fingers of the cathode. Preferably, the anode and the cathode are separated by a dielectric gap of 0.4 φ micrometers to 2.0 micrometers wide. GAS 081.9 Preferably, the organometallic complex is an organic ruthenium complex. GAS081.10 Preferably, the LOC device also has an electrochemical co-reactant 〇GAS081.il that is present with the organometallic complex during electrochemiluminescence. Preferably, the CMOS circuit has a light sensor for use. Photons emitted by the organometallic complex are sensed. GAS081.12 Preferably, the LOC device also has a hybridization chamber -23-201211533 array, wherein each of the hybridization chambers has a pair of electrodes and contains a plurality of probes, and the nucleic acids of the probes in each of the hybridization chambers The sequence is different from the nucleic acid sequence in at least one other hybridization chamber in the array such that a plurality of target nucleic acid sequences can be detected. GAS 081.13 Preferably, the CMOS circuit is positioned between the hybridization chambers and the support substrate such that the light sensor is adjacent to the hybridization chambers. Preferably, the light sensor is an array of photodiodes, the photodiode array being positioned such that each of the photodiodes corresponds to one of the hybridization chambers. GAS081.15 Preferably, the photodiodes have planar active surface regions for receiving light from the organometallic complex, each of the active surface regions being in the same plane, and the electrodes are one layer Patterned to form a separate conductive material of the anode and cathode, the plane of the layer being parallel to the plane of the active surface region of the photodiodes 〇GAS08 1.16, preferably, each of the electrodes is centered An electrode system causes oxidation or reduction of the organometallic complex to produce a working electrode of an excited species that emits photons, the working electrode being positioned such that the probes are interposed between the photodiode and the working electrode between. GAS081.17 Preferably, the photodiodes have a planar active surface region for receiving light from the organometallic complex, and the working electrode has optical coupling with an active surface region of the photodiode The surface region, the working electrode is configured such that the optically coupled surface area is greater than 50% of the active surface area of the photodiode. Preferably, the LOC device also has a polymerase interlocking reaction (PCR) portion for amplifying the target nucleic acid sequence in the sample. GAS081.19 Preferably, the PCR portion has a heater element for heating the nucleic acid sequence circulating the target and a polymerase, the heater element being configured to be operationally controlled by the C Ο S circuit. GAS 08 1.20 Preferably, the LOC device also has a plurality of sensors coupled to the CMOS circuit for feedback control of the electrodes and the heater elements. Electrochemiluminescence has the advantage of producing effective light at controlled locations in a microfluidic environment. In addition, synchronizing with the sensor is more convenient than a technology such as fluorescent. This LOC device has the benefit of less complex design and manufacturing requirements and therefore results in a simpler, more reliable manufacturing. The advantage of this chemistry is that the reporter complex is not consumed during photon emission, which increases the signal level for a given reporter concentration. GAS082.1 This aspect of the invention provides a wafer-on-lab (LOC) device for detecting a target nucleic acid sequence in a sample, the LOC device comprising: a support substrate; having complementary to the target nucleic acid sequence for use in forming a probe a nucleic acid sequence of a needle-target hybrid and a probe of an organic ruthenium complex: an electrode for generating an excited state of an organic ruthenium complex, wherein the organic ruthenium complex emits photons of light in the excited state; A CMOS circuit that applies a voltage to the electrodes. -25- 201211533 GAS082.2 Preferably, each of the probes has a functional portion that is resonated by resonance energy to quench photon emission from the organic germanium complex. GAS 082.3 Preferably, the probe is configured such that when the probe forms a probe-target hybrid, the functional moiety for quenching photon emission from the organic ruthenium complex is further derived from The organic germanium complex 〇GAS082.4 preferably, the CMOS circuit is configured to provide electrical pulses to the electrodes. GAS 08 2.5 Preferably, the electrical pulse has a period of less than 0.69 seconds. GAS 082.6 Preferably, the electrical current of the electrical pulse is between 0.1 nanoamperes and 69.0 nanoamperes. GAS 082.7 Preferably, the electrodes have an anode and a cathode, each having a finger configuration, the finger structures being configured to intersect the fingers of the anode with the fingers of the cathode. GAS 082.8 Preferably, the anode and cathode are separated by a dielectric gap of between 0.4 microns and 2.0 microns. GAS 082.9 Preferably, the light sensor has a planar active surface area for receiving light from the organic germanium complex and the electrodes have a thickness in the direction perpendicular to the active surface area of more than 2 microns. GAS 08 2.1 0 Preferably, the LOC device also has an electrochemical co-reactant with the organic ruthenium complex molecule during electrochemiluminescence. GAS082.il Preferably, the CMOS circuit has a light sensor for 201211533 for sensing photons emitted by the organic germanium complex molecule. GAS082.12 Preferably, the LOC device also has an array of hybridization chambers, wherein each of the hybridization chambers has a pair of electrodes and contains a plurality of probes, and the nucleic acid sequence of the probes in each of the hybridization chambers The nucleic acid sequences in at least one other hybridization chamber in the array are different such that a plurality of the subject nucleic acid sequences can be detected. GAS 082.1 3 Preferably, the CMOS circuit is positioned between the hybridization φ chamber and the support substrate such that the photosensor is adjacent to the hybridization chambers. Preferably, the light sensor is an array of photodiodes, the photodiode array being positioned such that each of the photodiodes corresponds to one of the hybridization chambers. GAS082.1 5 Preferably, the photodiodes have planar active surface regions for receiving light from the organic germanium complex, each of the active surface regions being in the same plane, and the electrodes are one layer Patterned to form the conductive material of the separate anode and cathode, the plane of the layer φ being parallel to the plane of the active surface region of the photodiodes 〇GAS082.1 6 preferably, each of these One of the electrode pairs causes oxidation or reduction of the organic ruthenium complex molecule to produce a working electrode of an excited species that emits photons, the working electrode being positioned such that the probes are interposed between the photodiodes Between the body and the working electrode. GAS 08 2.1 7 Preferably, the photodiodes have a planar active surface region for receiving light from the organic germanium complex molecule, and the working electrode has an active surface region light with the photodiode Coupling Table • 27-201211533 Surface area, the working electrode is configured such that the optically coupled surface area is greater than 50% of the active surface area of the photodiode. GAS08 2.1 8 Preferably, the LOC device also has a polymerase interlocking reaction (PC R) portion for amplifying the target nucleic acid sequence in the sample. GAS 082.1 9 Preferably, the PCR portion has a heater element for heating the nucleic acid sequence circulating the target and a polymerase, the heater element being configured to be operationally controlled by the CMOS circuit. GAS 082.20 Preferably, the LOC device also has a plurality of sensors coupled to the CMOS circuit for feedback control of the electrodes and the heater elements. Electrochemiluminescence has the advantage of producing effective light at controlled locations in a microfluidic environment. In addition, synchronizing with the sensor is more convenient than a technology such as fluorescent. This enables more sensitive and specific detection of the target DNA. This LOC device has the benefit of less complex design and manufacturing requirements and will therefore result in a simpler, more reliable manufacturing. The benefit of this chemistry is that the reporter complex is not consumed during photon emission, which increases the signal level for a given reporter concentration. GAS0 8 3.1 This aspect of the invention provides a wafer-on-lab (LOC) device for detecting a target nucleic acid sequence in a sample, the LOC device comprising: having a nucleic acid sequence complementary to the target for forming a probe-target a nucleic acid sequence of a hybrid and a probe of an electrochemiluminescence (ECL) luminophore; an electrode for generating an excited state of an ECL luminophore, wherein the ECL luminophore emits photons of light in the excited state of -28-201211533; An electrochemical co-reactant present with the luminophore during electrochemiluminescence. GAS 08 3.2 Preferably, each of the probes has a functional portion that is resonated by resonance energy to quench photon emission from the ECL luminophore. GAS083.3 Preferably, the probe is configured such that when the probe forms a probe-target hybrid, the functional portion for quenching photon emission from the ECL luminophore φ is further derived from The ECL luminophore. GAS 08 3.4 Preferably, the LOC device also has a CMOS circuit configured to provide electrical pulses to the electrodes. GAS08 3.5 Preferably, the electrical pulse has a period of less than 0.69 seconds. Preferably, the electrical current of the electrical pulse is between 0.1 nanoamperes and 69.0 nanoamperes. GAS 08 3.7 Preferably, the electrodes have an anode and a cathode, each having a finger-like configuration, the finger structures being configured to intersect the fingers of the anode with the fingers of the cathode. GAS 08 3.8 Preferably, the anode and cathode are separated by a dielectric gap of 0.4 microns to 2.0 microns. • GAS 083.9 Preferably, the luminophore is an organometallic complex. GAS 08 3.10 Preferably, the organometallic complex is an organogermanium complex molecule. GAS 083.1 1 Preferably, the CMOS circuit has a light sensor for sensing photons emitted by the ECL luminophore. -29-201211533 GAS083.1 2 Preferably, the LOC device also has an array of hybridization chambers, wherein each of the hybridization chambers has a pair of electrodes and contains a plurality of probes, the probes in each of the hybridization chambers The nucleic acid sequence is different from the nucleic acid sequence in at least one other hybridization chamber in the array such that a plurality of target nucleic acid sequences can be detected. Preferably, the LOC device also has a support substrate, wherein the CMOS circuit is located between the hybridization chamber and the support substrate such that the light sensor is adjacent to the hybrid chambers GAS083.1 4 Preferably, the light sensor is an array of photodiodes, the position of the photodiode array being configured such that each of the photodiodes corresponds to one of the hybridization chambers. GAS08 3.1 5 Preferably, the photodiodes have planar active surface regions for receiving light from the luminophore, each of the active surface regions being in the same plane, and the layers of the electrodes are patterned ( To form a conductive material of the separate anode and cathode, the plane of the layer being parallel to the plane of the active surface region of the photodiodes. GAS08 3.1 6 Preferably, one of the electrode pairs A working electrode that causes oxidation or reduction of the luminophore to produce an excited species that emits photons, the working electrode being positioned such that the probes are interposed between the photodiode and the working electrode. GAS 08 3.1 7 Preferably, the photodiodes have a planar active surface region for receiving light from the luminophore, and the working electrode has a surface region optically coupled to an active surface region of the photodiode The 8-30-201211533 working electrode is configured such that the optically coupled surface area is greater than 50% of the active surface area of the photodiode. GAS 083.1 8 Preferably, the LOC device also has a polymerase-associated reaction (PCR) portion for amplifying the target nucleic acid sequence in the sample.

GAS 083.19 Preferably, the PCR portion has a heater element for heating the nucleic acid sequence circulating the target and a polymerase, the heater element being configured to be operationally controlled by the CMOS circuit. GAS 083.20 Preferably, the LOC device also has a plurality of sensors coupled to the CMOS circuit for feedback control of the electrodes and the heater elements. Electrochemiluminescence has the advantage of producing effective light at controlled locations in a microfluidic environment. In addition, synchronizing with the sensor is more convenient than a technology such as fluorescent. This enables more sensitive and specific detection of the target DNA. This LOC device has the benefit of less complex design and manufacturing requirements and will therefore result in a simpler, more reliable manufacturing. The use of a co-reactant increases the signal intensity and reduces the voltage required to produce the electroluminescent signal. GAS084.1 This aspect of the invention provides a wafer-on-lab (LOC) device for detecting a target nucleic acid sequence in a sample, the LOC device comprising: a support substrate; having a complement to the target nucleic acid sequence for use in forming a probe a nucleic acid sequence of a needle-target hybrid and a probe of an electrochemiluminescence (ECL) luminophore; an electrode for generating an excited state of an ECL luminophore, wherein the EC L luminophore emits light in the excited state -31 - 201211533 a photon; and a CMOS circuit between the support substrate and the probes for applying a voltage to the electrodes. GAS 084.2 Preferably, each of the probes has a functional portion that is resonated by resonance energy to quench photon emission from the ECL luminophore. Preferably, the probe is configured such that when the probe forms a probe-target hybrid, the functional portion for quenching photon emission from the ECL luminophore is further derived from the ECL luminophore. G A S 0 8 4.4 Preferably, the C Μ Ο S circuit is configured to provide electrical pulses to the electrodes. GAS 084.5 Preferably, the electrical pulse has a period of less than 0.69 seconds. GAS 084.6 Preferably, the electrical current of the electrical pulse is between 0.1 nanoamperes and 69.0 nanoamperes. GAS 084.7 Preferably, the electrodes have an anode and a cathode, each having a finger configuration, the finger structures being configured to intersect the fingers of the anode with the fingers of the cathode. GAS 084.8 Preferably, the anode and cathode are separated by a dielectric gap of 0.4 microns to 2.0 microns. GAS 084.9 Preferably, the luminophore is an organometallic complex. GAS 084.1 0 Preferably, the organometallic complex is an organogermanium complex molecule. GAS 084. il. Preferably, the CMOS circuit has a light sensor for sensing photons emitted by the ECL luminophore. -32- 201211533 GAS084.1 2 Preferably, the LOC device also has an array of hybridization chambers, wherein each of the hybridization chambers has a pair of electrodes and contains a plurality of probes, the probes in each of the hybridization chambers The nucleic acid sequence is different from the nucleic acid sequence in at least one other hybridization chamber in the array such that a plurality of target nucleic acid sequences can be detected. GAS 084.1 3 Preferably, the CMOS circuit is located between the hybridization chambers and the support substrate such that the light sensors are adjacent to the hybridization chambers. GAS 084.14 Preferably, the light sensor is an array of photodiodes, the photodiode array being configured such that each of the photodiodes corresponds to one of the hybridization chambers. GAS 084.1 5 Preferably, the photodiodes have planar active surface regions for receiving light from the luminophore, each of the active surface regions being in the same plane, and the layers of the electrodes being patterned ( Patterned to form a conductive material of the separate anode and cathode, the plane of the layer being parallel to the plane of the active surface area of the photodiodes. GAS 084.1 6 Preferably, one of each of the pair of electrodes The electrode system causes oxidation or reduction of the luminophore to produce a working electrode of an excited species that emits photons, the working electrode being positioned such that the probes are interposed between the photodiode and the working electrode. GAS 084.1 7 Preferably, the photodiodes have a planar active surface area for receiving light from the luminophore, and the working electrode has a surface area optically coupled to an active surface area of the photodiode The working electrode is configured such that the surface area of the optical coupling is greater than 50% of the active surface area of the photo-33-201211533 diode. GAS 084.1 8 Preferably, the LOC device also has a polymerase linkage reaction (PCR) portion for amplifying the target nucleic acid sequence in the sample. GAS 084.1 9 Preferably, the PCR portion has a heater element for heating the nucleic acid sequence circulating the target and a polymerase, the heater element being configured to be operationally controlled by the CMOS circuit. GAS 084.20 Preferably, the LOC device also has a plurality of sensors coupled to the CMOS circuit for feedback control of the electrodes and the heater elements. Electrochemiluminescence has the advantage of producing effective light at controlled locations in a microfluidic environment. In addition, synchronizing with the sensor is more convenient than a technology such as fluorescent. This enables more sensitive and specific detection of the target DNA. This LOC device has the benefit of less complex design and manufacturing requirements and will therefore result in a simpler, more reliable manufacturing. This increases the specificity of the target molecule. GAS08 5.1 This aspect of the invention provides a wafer-on-lab (LOC) device for detecting a target nucleic acid sequence in a sample, the LOC device comprising: a support substrate; having a nucleic acid sequence complementary to the target for forming a probe a nucleic acid sequence of the target hybrid, an electrochemiluminescence (ECL) luminophore, and a probe for quenching a functional portion of photon emission from the ECL luminophore by resonance energy transfer; -34- 201211533 for generating EC L An electrode of an excited state of the luminophore 'in the excited state, the ECL luminophore emits photons of light; and a CMOS circuit interposed between the support substrate and the probes for applying a voltage to the electrodes. GAS 085.2 Preferably, the LOC device also has an electrochemical co-reactant present with the luminophore during electrochemiluminescence. GAS 08 5.3 Preferably, the probe is configured such that when the probe forms a probe-target hybrid, the functional portion for quenching photon emission from the ECL luminophore is further derived from the ECL Luminance* GAS 0 8 5.4 Preferably, the CMOS circuit is configured to provide electrical pulses to the electrodes. Preferably, the electrical pulse has a period of less than 0.69 seconds, as in the patented LOC device GAS 08 5.5. GAS 08 5.6 Preferably, the electrical current of the electrical pulse is between 0.1 nanoamperes and 69.0 nanoamperes. GAS 08 5.7 Preferably, the electrodes have an anode and a cathode, each having a finger configuration, the finger structures being configured to intersect the fingers of the anode with the fingers of the cathode. GAS 085.8 Preferably, the anode and cathode are separated by a dielectric gap of 0.4 microns to 2 microns wide. GAS08 5.9 Preferably, the luminophore is an organometallic complex. GAS 085.1 0 Preferably, the organometallic complex is an organogermanium complex molecule. GAS085.il Preferably, the CMOS circuit has a light sensor for sensing photons emitted by the ECL luminophore with -35-201211533. GAS 08 5.1 2 Preferably, the LOC device also has an array of hybridization chambers, wherein each of the hybridization chambers has a pair of electrodes and contains a plurality of probes, and the nucleic acid sequence of the probes in each of the hybridization chambers The nucleic acid sequences in at least one other hybridization chamber in the array are different such that a plurality of target nucleic acid sequences can be detected. GAS085.13 Preferably, the CMOS circuit is positioned between the hybridization chambers and the support substrate such that the light sensor is adjacent to the hybridization chambers. GAS 08 5.1 4 Preferably, the light sensor is an array of photodiodes, the position of the photodiode array being configured such that each of the photodiodes respectively corresponds to one of the hybrid chambers . GAS 08 5.1 5 Preferably, the photodiodes have planar active surface regions for receiving light from the luminophore, each of the active surface regions being in the same plane, and the layers of the electrodes are patterned To form a conductive material of the separate anode and cathode, the plane of the layer being parallel to the plane of the active surface area of the photodiode. GAS 08 5.1 6 Preferably, one of each of the pair of electrodes The electrode system causes oxidation or reduction of the luminophore to produce a working electrode of an excited species that emits photons, the working electrode being positioned such that the probes are interposed between the photodiode and the working electrode. GAS 08 5.1 7 Preferably, the photodiodes have a planar active surface area for receiving light from the luminophore, and the working electrode has a surface area optically coupled to an active surface area of the photodiode The -36-8 201211533 working electrode is configured such that the optically coupled surface area is greater than 50% of the active surface area of the photodiode. GAS 085.1 8 Preferably, the LOC device also has a polymerase chain reaction (PCR) portion for amplifying the target nucleic acid sequence in the sample. GAS 08 5.1 9 Preferably, the PCR portion has a heater element for heating the nucleic acid sequence of the target and a polymerase, the heater element φ being configured to be operationally controlled by the C Ο S circuit. GAS 08 5.20 Preferably, the LOC device also has a plurality of sensors coupled to the CMOS circuit for feedback control of the electrodes and the heater elements. Electrochemiluminescence has the advantage of producing effective light at controlled locations in a microfluidic environment. In addition, synchronizing with the sensor is more convenient than a technology such as fluorescent. This enables more sensitive and specific detection of the target DNA. This LOC device has the benefit of less complex design and manufacturing requirements, and φ will therefore result in a simpler, more reliable manufacturing. This signal change based on complementarity provides the advantage of obtaining a consistently formatted signal. There is no need to clean, otherwise sensitize or develop the steps to produce a signal whose intensity changes with the presence of the target. GAS086.1 This aspect of the invention provides a wafer-on-lab (LOC) device for detecting a target nucleic acid sequence in a sample, the LOC device comprising: having a nucleic acid sequence complementary to the target for forming a probe-target a nucleic acid sequence of a hybrid, an electrochemiluminescence (ECL) luminophore, and a probe that is capable of quenching a functional portion of photon emission from the ECL luminophore by resonance energy-37-201211533; and for generating ECL luminescence An electrode of an excited state of the cluster, in which the ECL luminophore emits photons of light; wherein during use, when a probe-target hybrid is formed, the functional moiety changes in proximity to the luminophore. GAS 086.2 Preferably, the LOC device also has an electrochemical co-reactant present with the luminophore during electrochemiluminescence. GAS086.3 Preferably, the probe is configured such that when the probe forms a probe-target hybrid, the functional moiety for quenching photon emission from the ECL luminophore is further derived from the ECL luminophore. GAS 086.4 Preferably, the LOC device also has a CMOS circuit configured to provide electrical pulses to the electrodes. GAS 0 8 6.5 Preferably, the electrical pulse has a period of less than 0.69 seconds. Preferably, the electrical current of the electrical pulse is between 0.1 nanoamperes and 69.0 nanoamperes. GAS 086.7 Preferably, the electrodes have an anode and a cathode, each having a finger configuration, the finger structures being configured to intersect the fingers of the anode with the fingers of the cathode. GAS 086.8 Preferably, the anode and cathode are separated by a dielectric gap of 4 micrometers to 2 micrometers wide. GAS 086.9 Preferably, the luminophore is an organometallic complex. GAS086.1 0 Preferably, the organometallic complex is an organic --38-201211533 conjugate molecule. GAS 086.11 Preferably, the CMOS circuit has a light sensor for sensing photons emitted by the ECL luminophore. GAS 08 6.1 2 Preferably, the LOC device also has an array of hybridization chambers, wherein each of the hybridization chambers has a pair of electrodes and contains a plurality of probes, and the nucleic acid sequence of the probes in each of the hybridization chambers The nucleic acid sequences in at least one other hybridization chamber in the array are different such that a plurality of target nucleic acid sequences can be detected. GAS 086.1 3 Preferably, the LOC device also has a support substrate, wherein the CMOS circuit is positioned to be interposed between the hybridization chamber and the support substrate such that the light sensor is adjacent to the hybridization chamber . GAS086.1 4 Preferably, the light sensor is an array of photodiodes, the position of the photodiode array being configured such that each of the photodiodes respectively corresponds to one of the hybrid chambers . GAS086.1 5 Preferably, the photodiodes have planar active surface regions for receiving φ light from the luminophore, each of the active surface regions being in the same plane, and the electrodes are patterned by a layer Patterned to form the conductive material of the separate anode and cathode, the plane of the layer being parallel to the plane of the active surface region of the photodiode 〇GAS086.1 6 Preferably, each of the electrodes is centered One of the electrodes causes oxidation or reduction of the luminophore to produce a working electrode of an excited species that emits photons, the working electrode being positioned such that the probes are interposed between the photodiode and the working electrode . -39- 201211533 GAS 08 6.1 7 Preferably, the photodiodes have a planar active surface area for receiving light from the luminophore, and the working electrode has an active surface area light with the photodiode The coupled surface region is configured such that the optically coupled surface region is greater than 50% of the active surface region of the photodiode. GAS 086.1 8 Preferably, the LOC device also has a polymerase linkage reaction (PCR) portion for amplifying the target nucleic acid sequence in the sample. GAS 08 6.1 9 Preferably, the PCR portion has a heater element for heating the nucleic acid sequence circulating the target and a polymerase, the heater element being configured to be operationally controlled by the CMOS circuit. GAS 08 6.20 Preferably, the LOC device also has a plurality of sensors coupled to the CMOS circuit for feedback control of the electrodes and the heater elements. Electrochemiluminescence has the advantage of producing effective light at controlled locations in a microfluidic environment. In addition, synchronizing with the sensor is more convenient than a technology such as fluorescent. This enables more sensitive and specific detection of the target DNA. This LOC device has the benefit of less complex design and manufacturing requirements and will therefore result in a simpler, more reliable manufacturing. This increases the specificity of the target molecule. The advantage of this signal change based on complementarity is the ability to obtain signals in a consistent format. There is no need to clean, additional sensitization or development steps to produce a signal whose intensity changes as the target is present. GAS087.1 This aspect of the invention provides a wafer-on-lab (L0C) device for detecting a target nucleic acid sequence in a sample, the LOC assembly 201211533 comprising: having a nucleic acid sequence complementary to the target for forming a probe a nucleic acid sequence of the target hybrid and a probe of an electrochemiluminescence (ECL) luminophore; an electrode for generating an excited state of the ECL luminophore, in which the photon of the ECL luminophore emits light; and a hybrid chamber An array, each of the hybridization chambers having a pair of electrodes and comprising a plurality of probes, wherein the nucleic acid sequence of the probes in each of the hybridization chambers is different from the nucleic acid sequence in at least one other hybridization chamber of the array of φ Multiple target nucleic acid sequences can be detected. GAS 087.2 Preferably, each of the probes has a functional portion that is resonated by resonance energy to quench photon emission from the ECL luminophore. GAS 087.3 Preferably, the probe is configured such that when the probe forms a probe-target hybrid, the functional portion for quenching photon emission from the ECL luminophore is further derived from the ECL luminophore. GAS 087.4 Preferably, the LOC device also has a CMOS circuit configured to provide electrical pulses to the electrodes. GAS 087.5 Preferably, the electrical pulse has a period of less than 0.69 seconds. Preferably, the electrical current of the electrical pulse is between 〇·1 nanoamperes to 69.0 nanoamperes. GAS 087.7 Preferably, the electrodes have an anode and a cathode, each having a finger configuration, the finger structures being configured to intersect the fingers of the anode with the fingers of the cathode. GAS 08 7.8 Preferably, the anode and cathode are separated by a dielectric gap of 0 -4 -41 - 201211533 microns to 2 microns wide. GAS 08 7.9 Preferably, the luminophore is an organometallic complex. GAS 08 7.1 0 Preferably, the organometallic complex is an organogermanium complex molecule. GAS 087.il Preferably, the CMOS circuit has a light sensor for sensing photons emitted by the ECL luminophore. GAS 08 7.1 2 Preferably, the LOC device also has an electrochemical co-reactant present with the luminophore during electrochemiluminescence. GAS 08 7.1 3 Preferably, the LOC device also has a support substrate, wherein the CMOS circuit is located between the hybridization chamber and the support substrate such that the light sensor is adjacent to the hybridization chambers. GAS 08 7.1 4 Preferably, the light sensor is an array of photodiodes, the position of the photodiode array being configured such that each of the photodiodes respectively corresponds to one of the hybrid chambers . GAS087.1 5 Preferably, the photodiodes have planar active surface regions for receiving light from the luminophore, each of the active surface regions being in the same plane, and the layers of the electrodes are patterned Patterned to form the conductive material of the separate anode and cathode, the plane of the layer being parallel to the plane of the active surface region of the photodiodes 〇GAS 08 7.1 6 Preferably, each of the electrodes is An electrode system causes oxidation or reduction of the luminophore to produce a working electrode of an excited species that emits photons, the working electrode being positioned such that the probes are interposed between the photodiode and the working electrode. -42- 201211533 GAS08 7.1 7 Preferably, the photodiodes have a planar active surface region for receiving light from the luminophore, and the working electrode has optical coupling with an active surface region of the photodiode The surface area of the working electrode is configured such that the optically coupled surface area is greater than 50% of the active surface area of the photodiode. GAS 08 7.1 8 Preferably, the LOC device also has a polymerase interlocking reaction (PCR) portion for amplifying the target nucleic acid sequence in the sample. GAS 087.1 9 Preferably, the PCR portion has a heater element for heating the nucleic acid sequence that circulates the target and a polymerase, the heater element being configured to be operationally controlled by the CMOS circuit. GAS 08 7.20 Preferably, the LOC device also has a plurality of sensors coupled to the CMOS circuit for feedback control of the electrodes and the heater elements. Electrochemiluminescence has the advantage of producing effective φ light at controlled locations in a microfluidic environment. In addition, synchronizing with the sensor is more convenient than a technology such as fluorescent. This enables more sensitive and specific detection of the target DNA. This LOC device has the benefit of less complex design and manufacturing requirements and will therefore result in a simpler, more reliable manufacturing. This increases the specificity of the target molecule. The LOC device design with parallel reaction sites allows parallel diagnostic testing to be performed in very small sample volumes, which increases the range and quality of diagnostic data obtained. GAS 088.1 This aspect of the invention provides a wafer-on-lab (LOC) device for detecting a target nucleic acid sequence in a sample, the LOC-43-201211533 comprising: having a nucleic acid sequence complementary to the target for formation a probe-target hybrid nucleic acid sequence and a probe for an electrochemiluminescence (ECL) luminophore; an electrode for generating an excited state of an ECL luminophore, in which the ECL luminophore emits photons of light; Light sensor 〇GAS088.2 for sensing photons emitted from the ECL luminophore. Preferably, each of the probes has a functional portion that is resonated by resonance energy transfer to quench photon emission from the ECL luminophore. GAS08 8.3 Preferably, the probe is configured such that when the probe forms a probe-target hybrid, the functional moiety for quenching photon emission from the ECL luminophore is further derived from the ECL Luminous group. GAS 088.4 Preferably, the LOC device also has a CMOS circuit configured to provide electrical pulses to the electrodes. GAS08 8.5 Preferably, the electrical pulse has a period of less than 0.69 seconds. GAS 08 8.6 Preferably, the electrical current of the electrical pulse is between 0.1 nanoamperes and 69.0 nanoamperes. GAS 08 8.7 Preferably, the electrodes have an anode and a cathode, each having a finger configuration, the finger structures being configured to intersect the fingers of the anode with the fingers of the cathode. GAS08 8.8 Preferably, the anode and cathode are separated by a dielectric gap of 0.4 microns to 2 microns wide. GAS 088.9 Preferably, the luminophore is an organometallic complex. -44 - 201211533 GAS08 8.1 0 Preferably, the organometallic complex is an organogermanium complex molecule. GAS088.il Preferably, the CMOS circuit is provided with the light sensor such that the light sensor is in close proximity to the hybrid chambers. GAS08 8.1 2 Preferably, the LOC device also has an array of hybridization chambers, wherein each of the hybridization chambers has a pair of electrodes and contains a plurality of probes, and the nucleic acid sequence of the probes in each of the hybridization chambers φ in the array differs in the nucleic acid sequence in at least one other hybridization chamber such that a plurality of target nucleic acid sequences can be detected. GAS 08 8.1 3 Preferably, the LOC device also has a support substrate, wherein the CMOS circuit is located between the hybridization chamber and the support substrate such that the light sensor is adjacent to the hybridization chambers. GAS 088.14 Preferably, the light sensor is an array of photodiodes, the positions of the photodiode array being configured such that each of the photodiodes corresponds to one of the hybridization chambers. φ GAS 088.1 5 Preferably, the photodiodes have planar active surface regions for receiving light from the luminophore, each of the active surface regions being in the same plane, and the layers of the electrodes are patterned Patterned to form the conductive material of the separate anode and cathode, the plane of the layer being parallel to the plane of the active surface region of the photodiodes 〇GAS 08 8.1 6 Preferably, each of the electrodes is An electrode system causes oxidation or reduction of the luminophore to produce a working electrode of an excited species that emits photons, the working electrode being positioned such that the probes are between -45 and 201211533 and the photodiode is associated with the work Between the electrodes. GAS 08 8.1 7 Preferably, the photodiodes have a planar active surface area for receiving light from the luminophore, and the working electrode has a surface area optically coupled to an active surface area of the photodiode The working electrode is configured such that the optically coupled surface area is greater than 50% of the active surface area of the photodiode. GAS 08 8.1 8 Preferably, the LOC device also has a polymerase linkage reaction (PCR) portion for amplifying the target nucleic acid sequence in the sample. GAS 088.1 9 Preferably, the PCR portion has a heater element for heating the nucleic acid sequence circulating the target and a polymerase, the heater element being configured to be operationally controlled by the C Μ O S circuit. GAS 08 8.20 Preferably, the LOC device also has a plurality of sensors coupled to the CMOS circuit for feedback control of the electrodes and the heater elements. The integrated light sensor has the advantage of higher light efficiency than the off-chip sensor design. Integrated light sensors have the advantage of being easier to synchronize with other system events. Integrated light sensors have the advantage of reducing the number of discrete components. Electrochemiluminescence has the advantage of producing effective light at controlled locations in a microfluidic environment. In addition, synchronizing with the sensor is more convenient than a technology system such as fluorescent. This LOC device has the benefit of less complex design and manufacturing requirements and therefore results in a simpler, more reliable manufacturing. This enables more sensitive and specific detection of the target DNA. GAS 08 9. 1 This aspect of the invention provides a wafer-on-lab (LOC) device for detecting a nucleic acid sequence in a sample of -46 - 201211533, the LOC device comprising: having a nucleic acid sequence complementary to the target a probe for forming a nucleic acid sequence of a probe-target hybrid and an electrochemiluminescence (ECL) luminophore; an electrode for generating an excited state of an ECL luminophore, wherein the ECL luminophore emits photons of light in the excited state; And a light sensor for sensing photons emitted from the ECL luminophore: wherein the electrodes are plates of a conductive material, the plates having peripheral edges that are optically coupled to the light sensor. GAS 089.2 Preferably, each of the probes has a functional portion that is resonated by resonance energy to quench photon emission from the ECL luminophore. GAS089.3 Preferably, the probe is configured such that when the probe forms a probe-target hybrid, the functional portion for quenching photon emission from the ECL luminophore is further derived from the ECL luminophore. GAS 089.4 Preferably, the LOC device also has a CMOS circuit configured to provide electrical pulses to the electrodes. GAS 089.5 Preferably, the electrical pulse has a period of less than 0.69 seconds. GAS 089.6 Preferably, the electrical current of the electrical pulse is between 0.1 nanoamperes and 69.0 nanoamperes. GAS 089.7 Preferably, the electrodes have an anode and a cathode, each having a finger configuration, the finger structures being configured to intersect the fingers of the anode with the fingers of the cathode. -47- 201211533 GAS089.8 Preferably, the anode and cathode are separated by a dielectric gap of 0.9 microns to 2 microns wide. GAS 089.9 Preferably, the luminophore is an organometallic complex. GAS 089.1 0 Preferably, the organometallic complex is an organogermanium complex molecule. GAS 089. il. Preferably, the CMOS circuit is provided with the light sensor such that the light sensor is in close proximity to the hybrid chambers. GAS089.1 2 Preferably, the LOC device also has an array of hybridization chambers, wherein each of the hybridization chambers has a pair of electrodes and contains a plurality of probes, and the nucleic acid sequence of the probes in each of the hybridization chambers The nucleic acid sequences in at least one other hybridization chamber in the array are different such that a plurality of target nucleic acid sequences can be detected. GAS 089.1 3 Preferably, the LOC device also has a support substrate, wherein the CMOS circuit is located between the hybridization chambers and the support substrate such that the light sensor is adjacent to the hybridization chambers. Preferably, the light sensor is an array of photodiodes, the photodiode array being positioned such that each of the photodiodes corresponds to one of the hybridization chambers. Preferably, the photodiodes have planar active surface regions for receiving light from the luminophore, each of the active surface regions being in the same plane, and the layers of the electrodes are patterned Patterned to form the conductive material of the separate anode and cathode, the plane of the layer being parallel to the plane of the active surface region of the photodiode. 8 48 - 201211533 GAS08 9.1 6 Preferably, each of the electrode pairs One of the electrodes causes oxidation or reduction of the luminophore to produce a working electrode of an excited species that emits photons, the working electrode being positioned such that the probes are interposed between the photodiode and the working electrode between. GAS089.1 7 Preferably, the photodiodes have a planar active surface region for receiving light from the luminophore, and the working electrode has a surface region optically coupled to an active surface region of the photodiode The working electrode is configured such that the optically coupled surface area is greater than 50% of the active surface area of the photodiode. GAS 08 9.1 8 Preferably, the LOC device also has a polymerase interlocking reaction (PCR) portion for amplifying the target nucleic acid sequence in the sample. GAS 089.1 9 Preferably, the PCR portion has a heater element for heating the nucleic acid sequence circulating the target and a polymerase, the heater element being configured to be operationally controlled by the C Ο S circuit. GAS089.20 Preferably, the LOC device also has a plurality of sensors coupled to the CMOS circuit for feedback control of the electrodes and the heater elements. The integrated light sensor has the advantage of higher light efficiency than the off-chip sensor design. Integrated light sensors have the advantage of being easier to synchronize with other system events. Integrated light sensors have the advantage of reducing the number of discrete components. Electrochemiluminescence has the advantage of producing effective light at controlled locations in a microfluidic environment. In addition, synchronizing with the sensor is more convenient than a technology system such as fluorescent. This enables more sensitive and specific detection of the target -49-201211533 DNA. This LOC device has the advantage of less complex design and manufacturing requirements and therefore results in a simpler, more reliable manufacturing. This LOC device design has the advantage of increasing the coupling between the light emitting region and the light sensor. GAS090.1 This aspect of the invention provides a wafer-on-lab (LOC) device for detecting a target nucleic acid sequence in a sample, the LOC device comprising: having a nucleic acid sequence complementary to the target for forming a probe-target a nucleic acid sequence of a hybrid and a probe of an electrochemiluminescence (ECL) luminophore; an electrode for generating an excited state of an ECL luminophore in which the photon of the ECL luminophore emits light; and for sensing from a photon sensor of the photon emitted by the ECL luminophore: wherein the electrodes have at least one working electrode that causes oxidation or reduction of the luminophore to generate an excited species that emits photons, the working electrode being in close proximity to the photosensor . GAS 090.2 Preferably, each of the probes has a functional portion that is resonated by resonance energy to quench photon emission from the E C L luminophore. GAS090.3 Preferably, the probe is configured such that when the probe forms a probe-target hybrid, the functional moiety for quenching photon emission from the ECL luminophore is further derived from the ECL luminophore. GAS 090.4 Preferably the LOC device also has a CMOS circuit configured to provide electrical pulses to the electrodes. G A S 090.5 Preferably, the electrical pulse has a period of less than 0.69 seconds -50 - 201211533 GAS090.6 Preferably, the electrical current of the electrical pulse is between 0.1 nanoamperes and 69.0 nanoamperes. GAS 090.7 Preferably, the electrodes have an anode and a cathode, each having a finger configuration, the finger structures being configured to intersect the fingers of the anode with the fingers of the cathode. GAS 090.8 Preferably, the anode and cathode are separated by a dielectric gap of 0.4 microns to 2 microns wide. φ GAS090.9 Preferably, the luminophore is an organometallic complex. GAS 090.1 0 Preferably, the organometallic complex is an organogermanium complex molecule. GAS090.1 1 Preferably, the CMOS circuit has a light sensor for sensing photons emitted by the ECL luminophore. GAS090.12 Preferably, the LOC device also has an array of hybridization chambers, wherein each of the hybridization chambers has a pair of electrodes and contains a plurality of probes, and the nucleic acid sequence of the probes in each of the hybridization chambers φ in the array differs in the nucleic acid sequence in at least one other hybridization chamber such that a plurality of target nucleic acid sequences can be detected. GAS 090.1 3 Preferably, the LOC device also has a support substrate, wherein the CMOS circuit is located between the hybridization chamber and the support substrate such that the light sensor is adjacent to the hybridization chambers. Preferably, the light sensor is an array of photodiodes whose positions are configured such that each of the photodiodes corresponds to one of the hybrid chambers. GAS090.1 5 Preferably, the photodiodes have planar active surface regions for receiving -51 - 201211533 light from the luminophore, each of the active surface regions being in the same plane, and the electrode systems A layer is patterned to form the conductive material of the separate anode and cathode, the plane of the layer being parallel to the plane of the active surface area of the photodiode 〇GAS090.1 6 preferably, such work Electrodes are plates of electrically conductive material that define a series of finger-like configurations to increase the length of the peripheral edge of the plate that is optically coupled to the photosensor. GAS090.1 7 Preferably, the photodiodes have a planar active surface region for receiving light from the luminophore, and the working electrode has a surface region optically coupled to an active surface region of the photodiode The working electrode is configured such that the optically coupled surface area is greater than 50% of the active surface area of the photodiode. GAS 090.1 8 Preferably, the LOC device also has a polymerase ligation reaction (PCR) portion for amplifying the target nucleic acid sequence in the sample. GAS 090.1 9 Preferably, the PCR portion has a heater element for heating the nucleic acid sequence circulating the target and a polymerase, the heater element being configured to be operationally controlled by the CMOS circuit. GAS090.20 Preferably, the LOC device also has a plurality of sensors coupled to the CMOS circuit for feedback control of the electrodes and the heater elements. The integrated light sensor has the advantage of higher light efficiency than the off-chip sensor design. Integrated light sensor has easier event with other systems -52-

201211533 The advantages of synchronization. Integrated light sensors have the advantage of reducing discrete components. Electrochemiluminescence has the advantage of controlling positional light in a microfluidic environment. In addition, synchronization with the sensor is more convenient than, for example, an egg system. This enables more sensitive and specific detection of DNA. » This LOC device has less complex design and manufacturing advantages and therefore will result in a simpler, more reliable manufacturing. The LOC meter has a GAS091.1 that increases the coupling between the light-emitting region and the light sensor. This aspect of the invention provides a wafer-on-lab (LOC) device for in-situ calibration of a nucleic acid sequence. Including: an electrode having a nucleic acid sequence complementary to the target nucleic acid sequence for forming a probe-crossbody and an electrochemiluminescence (ECL) luminophore for generating an excited state of the ECL luminophore, in the state a photon emitted by the ECL luminophore; and a photon for sensing photons emitted from the ECL luminophore; wherein the photosensor has an active surface region for receiving the ECL luminophore from the surface and the electrodes are The thickness of the direction perpendicular to the active surface regions of the photodiodes H is between 0.25 micrometers and 23⁄4 GAS 091.2. Preferably, each of the probes has a common twisting to quench photon emission from the ECL luminophores. Functional 茜GAS091.3 Preferably, when the probe is configured to form a probe-target hybrid, the functional portion for quenching photon emission from the ECL is further derived from the ECL The number of sub-quantities Good means disposed seeking the advantages of the subject of measurement technology. Measuring sample LOC loading • Standard miscellaneous: excitation; sensor. Light flat: plane [m. : Energy turns ί points. The probe luminophore: group. -53- 201211533 GAS091.4 Preferably the LOC device also has a CMOS circuit configured to provide electrical pulses to the electrodes. GAS 091.5 Preferably, the electrical pulse has a period of less than 0.69 seconds. GAS091.6 Preferably, the electrical current of the electrical pulse is between 0.1 nanoamperes and 69.0 nanoamperes. GAS 091.7 Preferably, the electrodes have an anode and a cathode, each having a finger configuration, the finger structures being configured to intersect the fingers of the anode with the fingers of the cathode. GAS 091.8 Preferably, the anode and cathode are separated by a dielectric gap of 0.4 microns to 2 microns wide. GAS 09 1.9 Preferably, the luminophore is an organometallic complex. GAS 091.10 Preferably, the organometallic complex is an organogermanium complex molecule. GAS091.il Preferably, the CMOS circuit has a light sensor for sensing photons emitted by the ECL luminophore. GAS091.12 Preferably, the LOC device also has an array of hybridization chambers, wherein each of the hybridization chambers has a pair of electrodes and contains a plurality of probes, and the nucleic acid sequence of the probes in each of the hybridization chambers The nucleic acid sequences in at least one other hybridization chamber in the array are different such that a plurality of the subject nucleic acid sequences can be detected. GAS091.13 Preferably, the LOC device also has a support substrate 'where the CMOS circuit is located between the hybridization chamber and the support substrate such that the light sensor is adjacent to the hybridization chambers. -54-201211533 GAS091.14 Preferably, the light sensor is an array of photodiodes, the position of the photodiode array being configured such that each of the photodiodes respectively corresponds to a hybrid chamber One of them. GAS 09 1.1 5 Preferably, the photodiodes have planar active surface regions for receiving light from the luminophore, each of the active surface regions being in the same plane, and the layers of the electrodes are patterned Patterned to form the conductive material of the separate anode and cathode, the plane of the layer being parallel to the plane of the active surface region of the photodiodes 〇GAS091.16 Preferably, each of the electrodes is centered One of the electrodes causes oxidation or reduction of the luminophore to produce a working electrode of an excited species that emits photons, the working electrode being positioned such that the probes are interposed between the photodiode and the working electrode . GAS091.17 Preferably, the photodiodes have a planar active surface region for receiving light from the luminophore, and the working electrode has a surface optically coupled to an active surface region of the photodiode The working electrode is configured such that the optically coupled surface area is greater than 50% of the active surface area of the photodiode. GAS 091.18 Preferably, the LOC device also has a polymerase chain reaction (PCR) portion for amplifying the target nucleic acid sequence in the sample. GAS091.19 Preferably, the PCR portion has a heater element for heating the nucleic acid sequence that circulates the target and a polymerase, the heater element being configured to be operationally controlled by the C Μ 0 S circuit. -55- 201211533 GAS09 1 .20 Preferably, the LOC device also has a plurality of sensors coupled to the CMOS circuit for feedback control of the electrodes and the heater elements. The integrated light sensor has the advantage of higher light efficiency than the off-chip sensor design. Integrated light sensors have the advantage of being easier to synchronize with other system events. Integrated light sensors have the advantage of reducing the number of discrete components. Electrochemiluminescence has the advantage of producing effective light at controlled locations in a microfluidic environment. In addition, synchronizing with the sensor is more convenient than a technology system such as fluorescent. This enables more sensitive and specific detection of the target DNA. This LOC device has the advantage of less complex design and manufacturing requirements and therefore results in a simpler, more reliable manufacturing. This LOC device design has the advantage of increasing the coupling between the light emitting region and the light sensor. GAS092.1 This aspect of the invention provides a wafer-on-lab (LOC) device for detecting a target nucleic acid sequence in a sample, the LOC device comprising: having a nucleic acid sequence complementary to the target for forming a probe-target a nucleic acid sequence of a hybrid and a probe of an electrochemiluminescence (ECL) luminophore; an electrode for generating an excited state of an ECL luminophore in which the photon of the ECL luminophore emits light: and for sensing from A photon sensor of the photon emitted by the ECL luminophore: wherein the electrodes are plates of a conductive material, the edge characteristics of the plates being configured such that the perimeter edges of each of the plates are greater than 128 microns. GAS 092.2 Preferably, each of the probes has a functional portion that is shifted by resonance energy to -56 - 201211533 to quench photon emission from the ECL luminophore. GAS092.3 Preferably, the probe is configured such that when the probe forms a probe-target hybrid, the functional moiety for quenching photon emission from the ECL luminophore is further derived from the ECL luminophore. GAS 092.4 Preferably, the LOC device also has a CMOS circuit configured to provide electrical pulses to the electrodes. GAS 092.5 Preferably, the electrical pulse has a period of φ of less than 0.69 seconds. GAS092.6 Preferably, the current of the electrical pulse is between 0.1 nanoamperes and 69.0 nanoamperes. GAS 092.7 Preferably, the electrodes have an anode and a cathode, each having a finger configuration, the finger structures being configured to intersect the fingers of the anode with the fingers of the cathode. GAS092.8 Preferably, the anode and cathode are separated by a dielectric gap of 0.4 microns to 2 microns wide. φ GAS092.9 Preferably, the luminophore is an organometallic complex. GAS 092.1 0 Preferably, the organometallic complex is an organogermanium complex molecule. GAS092.il Preferably, the CMOS circuit has a light sensor for sensing photons emitted by the ECL luminophore. GAS092.12 Preferably, the LOC device also has an array of hybridization chambers, wherein each of the hybridization chambers has a pair of electrodes and contains a plurality of probes, and the nucleic acid sequence of the probes in each of the hybridization chambers The nucleic acid sequences in at least one other hybridization chamber in the array are different such that a plurality of the labeled -57-201211533 nucleic acid sequences can be detected. GAS 092.1 3 Preferably, the LOC device also has a support substrate, wherein the CMOS circuit is located between the hybridization chamber and the support substrate such that the light sensor is adjacent to the hybridization chambers. GAS092.1 4 Preferably, the light sensor is an array of photodiodes, the position of the photodiode array being configured such that each of the photodiodes respectively corresponds to one of the hybrid chambers . GAS092.1 5 Preferably, the photodiodes have planar active surface regions for receiving light from the luminophore, each of the active surface regions being in the same plane, and the layers of the electrodes are patterned Patterned to form the conductive material of the separate anode and cathode, the plane of the layer being parallel to the plane of the active surface region of the photodiodes 〇GAS092.1 6 preferably, each of the electrodes is An electrode system causes oxidation or reduction of the luminophore to produce a working electrode of an excited species that emits photons, the working electrode being positioned such that the probes are interposed between the photodiode and the working electrode. GAS092.1 7 Preferably, the photodiodes have a planar active surface area for receiving light from the luminophore, and the working electrode has a surface area optically coupled to an active surface area of the photodiode The working electrode is configured such that the optically coupled surface area is greater than 50% of the active surface area of the photodiode. GAS 092.1 8 Preferably, the LOC device also has a polymerase interlocking reaction (PCR) portion for amplifying the target nucleic acid sequence 201211533 column in the sample. GAS 092.1 9 Preferably, the PCR portion has a heater element for heating the nucleic acid sequence circulating the target and a polymerase, the heater element being configured to be operationally controlled by the C Ο S circuit. GAS 092.20 Preferably, the LOC device also has a plurality of sensors coupled to the CMOS circuit for feedback control of the electrodes and the heater elements. The φ integrated light sensor has the advantage of higher light efficiency 1 than the off-chip sensor design. Integrated light sensors have the advantage of being easier to synchronize with other system events. Integrated light sensors have the advantage of reducing the number of discrete components. Electrochemiluminescence has the advantage of producing effective light at controlled locations in a microfluidic environment. In addition, synchronizing with the sensor is more convenient than a technology system such as fluorescent. This enables more sensitive and specific detection of the target DNA. This LOC device has the advantage of less complex design and manufacturing requirements and therefore results in a simpler, more reliable manufacturing. The LOC device has an advantage of increasing the coupling between the light-emitting region and the light sensor. GAS093.1 This aspect of the invention provides a wafer-on-lab (for wafer-on-a-chip) for detecting a nucleic acid sequence in a sample ( a LOC device comprising: a probe having a nucleic acid sequence complementary to the target nucleic acid sequence for use in forming a probe-target hybrid and an electrochemiluminescence (ECL) luminophore; and for generating an ECL luminophore An excited state electrode in which the ECL luminophore emits photons of light; wherein the electrodes have an anode and a cathode, each anode and cathode having a finger-like configuration configured to cross each other -59-201211533. GAS 093.2 Preferably, each of the probes has a functional portion that is resonated by resonance energy to quench photon emission from the ECL luminophore. GAS093.3 Preferably, the probe is configured such that when the probe forms a probe-target hybrid, the functional moiety for quenching photon emission from the ECL luminophore is further derived from the ECL luminophore. GAS 093.4 Preferably, the LOC device also has a CMOS circuit configured to provide electrical pulses to the electrodes. GAS 093.5 Preferably, the electrical pulse has a period of less than 0.69 seconds. GAS093.6 Preferably, the electrical current of the electrical pulse is between 0.1 nanoamperes and 69.0 nanoamperes. GAS 093.7 Preferably, the LOC device also has an electrochemical co-reactant present with the ECL luminophore during electrochemiluminescence. GAS 093.8 Preferably, the anode and cathode are separated by a dielectric gap of 0.4 microns to 2 microns wide. GAS 093.9 Preferably, the luminophore is an organometallic complex. GAS 093.1 0 Preferably the organometallic complex is an organogermanium complex molecule. GAS093.il Preferably, the CMOS circuit has a light sensor for sensing photons emitted by the ECL luminophore. GAS093.1 2 Preferably, the LOC device also has an array of hybridization chambers, wherein each of the hybridization chambers has a pair of electrodes and contains a plurality of probes, and the nucleic acid sequence of the probes in each of the hybridization chambers The nucleic acid sequences in the at least one other hybridization chamber of -60-201211533 in the array are different such that a plurality of target nucleic acid sequences can be detected. GAS 093.1 3 Preferably, the LOC device also has a support substrate, wherein the CMOS circuit is located between the hybridization chamber and the support substrate such that the light sensor is adjacent to the hybridization chambers. GAS093.1 4 Preferably, the light sensor is an array of photodiodes, the position of the photodiode array being configured such that each of the photodiodes are respectively corresponding to one of the hybrid chambers By. GAS093.1 5 Preferably, the photodiodes have planar active surface regions for receiving light from the luminophore, each of the active surface regions being in the same plane, and the layers of the electrodes are patterned Patterned to form the conductive material of the separate anode and cathode, the plane of the layer being parallel to the plane of the active surface region of the photodiodes 〇GAS093.1 6 preferably, each of the electrodes is An electrode system φ causes oxidation or reduction of the luminophore to produce a working electrode of an excited species that emits photons, the working electrode being positioned such that the probes are interposed between the photodiode and the working electrode . GAS093.1 7 Preferably, the photodiodes have a planar active surface region for receiving light from the luminophore, and the working electrode has a surface region optically coupled to an active surface region of the photodiode The working electrode is configured such that the optically coupled surface area is greater than 50% of the active surface area of the photodiode. GAS 093.1 8 Preferably, the LOC device also has a polymerase ligature 201211533 lock reaction (PCR) portion for amplifying the target nucleic acid sequence in the sample. GAS 093.1 9 Preferably, the PCR portion has a heater element for heating the nucleic acid sequence circulating the target and a polymerase, the heater element being configured to be operationally controlled by the CMOS circuit. GAS093.20 Preferably, the LOC device also has a plurality of sensors coupled to the CMOS circuit for feedback control of the electrodes and the heater elements. The integrated light sensor has the advantage of higher light efficiency than the off-chip sensor design. Integrated light sensors have the advantage of being easier to synchronize with other system events. Integrated light sensors have the advantage of reducing the number of discrete components. Electrochemiluminescence has the advantage of producing effective light at controlled locations in a microfluidic environment. In addition, synchronizing with the sensor is more convenient than a technology system such as fluorescent. This enables more sensitive and specific detection of the target DNA. This LOC device has the advantage of less complex design and manufacturing requirements and therefore results in a simpler, more reliable manufacturing. This LOC device design has the advantage of increasing the coupling between the light emitting region and the light sensor. GAS094.1 This aspect of the invention provides a wafer-on-lab (LOC) device for detecting a target nucleic acid sequence in a sample, the LOC device comprising: having a nucleic acid sequence complementary to the target for forming a probe-target a nucleic acid sequence of a hybrid and a probe for an electrochemiluminescence (ECL) luminophore; an electrode pair, wherein one of each of the pair of electrodes causes oxidation or reduction of the luminophore to produce a working electrode of an excited species that emits photons -62-201211533: and a photodiode corresponding to each of the pair of electrodes; wherein the photodiodes have a planar active surface region for receiving light from the luminophore, and the working electrode has the optoelectronic The active surface region of the diode is optically coupled to the surface region, the working electrode being configured such that the optically coupled surface region is greater than 50% of the active surface region of the photodiode. φ GAS 094.2 Preferably, the two electrodes of one of the pairs of electrodes have a finger configuration configured to intersect each other. GAS 094.3 Preferably, each of the probes has a functional moiety for quenching photon emission from the ECL luminophore by resonance energy transfer. GAS094.4 Preferably, the probe is configured to When the probe forms a probe-target hybrid, the functional moiety for quenching photon emission from the ECL luminophore is further derived from the ECL luminophore. GAS 094.5 Preferably, the LOC device also has a CMOS circuit configured to provide electrical pulses to the electrodes. GAS 094.6 Preferably, the electrical pulse has a period of less than 0.69 seconds. GAS094.7 Preferably, the electrical current of the electrical pulse is between 0.1 nanoamperes and 69.0 nanoamperes. GAS 094.8 Preferably, the working electrode is configured such that the surface area of the optical coupling is greater than 90% of the active surface area of the photodiode. Preferably, GAS094.9 is between -63 and 201211533 between the electrodes of each of the pairs of electrodes separated by a dielectric gap of 0.4 microns to 2 microns. GAS094.10 Preferably, the luminophore is an organometallic complex. GAS094.il Preferably, the organometallic complex is an organogermanium complex molecule. GAS 094.1 2 Preferably, the CMOS circuit has a light sensor for sensing photons emitted by the ECL luminophore. GAS094.1 3 Preferably, the LOC device also has an array of hybridization chambers, wherein each of the hybridization chambers has one of the pair of electrodes and contains a plurality of probes, probes in each of the hybridization chambers The nucleic acid sequence is different from the nucleic acid sequence in at least one other hybridization chamber in the array such that a plurality of the target nucleic acid sequences can be detected. GAS 094.1 4 Preferably, the LOC device also has a support substrate, wherein the CMOS circuit is located between the hybridization chamber and the support substrate such that the photodiodes are adjacent to the hybridization chambers. GAS 094.1 5 Preferably, the working electrode is positioned such that the probes are interposed between the photodiode and the working electrode. GAS094.1 6 Preferably, each of the active surface regions is in the same plane, and the electrodes are patterned to form the conductive material of the separate anode and cathode, the plane of the layer and the layer The planes of the active surface regions of the photodiodes are parallel. Preferably, the electrodes are between 0.25 microns and 2 microns thick in the direction perpendicular to the planar active surface area of the photodiodes. GAS094.1 8 Preferably, the LOC device also has a polymerase-64-

201211533 Lock Reaction (PC R), which is used to amplify the label in this sample. GAS094.19 Preferably, the PCR portion has a heater element for adding the target nucleic acid sequence to a polymerase, the heater being configured to be operationally controlled by the C Ο S circuit. GAS094.20 Preferably, the LOC device also has multiple CMOS circuit-connected sensors for feedback control of the electrode elements. The integrated light sensor has the advantage of narrow efficiency compared to the off-chip sensor design. The integrated light sensor has the advantage of being easier to synchronize with other abandonment. Integrated light sensors have the advantage of reducing discrete components. Electrochemiluminescence has the advantage of controlling the potential light in a microfluidic environment. In addition, synchronization with the sensor is more convenient than for example. This allows for more sensitive and specific detection of DNA. This LOC device has less complex design and manufacturing considerations and will therefore result in a simpler, more reliable manufacturing. The LOC meter has a GAS095.1 that increases the coupling between the light-emitting region and the light sensor. This aspect of the invention provides a wafer-on-lab (LOC) device for the nucleic acid sequence of the target in the yang, the set comprising: A probe having a nucleic acid sequence complementary to the target nucleic acid sequence for forming a probe crossbody and an electrochemiluminescence (ECL) luminophore for generating an excited state of the ECL luminophore, in which the ECL luminophore emits Photon photon; where I nucleic acid sequence thermal cycle ^ component system, and the device and the "higher optical event: the number of ΐ ΐ 有 ΐ ΐ ΐ ΐ ΐ 5 5 5 ί ί ί ί ί ί The sample LOC is loaded with the standard: and [excited-65-201211533. The electrodes are arranged in pairs, and one of the electrode pairs causes oxidation or reduction of the luminophore to produce an excited species that emits photons. An electrode, the position of the working electrode being configured such that the probe is interposed between the photosensor and the working electrode. GAS 095.2 Preferably, each of the probes has quenching by resonance energy transfer Functional portion of the photon emission of the ECL luminophore. GAS 095.3 Preferably, the probe is configured to quench the elucent cluster from the ECL when the probe forms a probe-target hybrid The functional portion of the photon emission is further derived from the ECL luminophore. GAS 095.4 Preferably, the LOC device also has a CMOS circuit configured to provide electrical pulses to the electrodes. GAS 095.5 Preferably, the electrical pulse Having a period of less than 0.69 seconds GAS 095.6 Preferably, the electrical current of the electrical pulse is between 0.1 Nai and 69.0 Nai. GAS 095.7 Preferably, the electrode pairs have an anode and a cathode, and each anode and cathode have a It is configured to have a finger-like configuration that intersects each other. GAS 095.8 Preferably, the anode and cathode are separated by a dielectric gap of 0.4 micrometers to 2.0 micrometers wide. GAS 095.9 Preferably, the luminophore is an organometallic complex. Preferably, the organometallic complex is an organogermanium complex molecule. GAS 095.1 1 Preferably, the CMOS circuit has a light sensor for sensing emission by the ECL luminophore Photon. 8 -66- 201 211533 .GAS095.1 2 Preferably, the LOC device also has an array of hybridization chambers, wherein each of the hybridization chambers has one of the pair of electrodes and contains a plurality of probes in each of the hybridization chambers The nucleic acid sequence of the probe is different from the nucleic acid sequence in at least one other hybridization chamber in the array such that a plurality of the target nucleic acid sequences can be detected. GAS 095.1 3 Preferably, the LOC device also has a support substrate, wherein A CMOS circuit is located between the hybridization chambers and the support substrate φ such that the light sensor is adjacent to the hybridization chambers. GAS095.1 4 Preferably, the light sensor is an array of photodiodes, the position of the photodiode array being configured such that each of the photodiodes respectively corresponds to one of the hybrid chambers . GAS 09 5.1 5 Preferably, the photodiodes have planar active surface regions for receiving light from the luminophore, each of the active surface regions being in the same plane, and the layers of the electrodes are patterned Patterned to form the conductive material of the separate anode and cathode, the plane of the layer φ is parallel to the plane of the active surface region of the photodiodes. GAS095.1 6 Preferably, the LOC device also has An electrochemical co-reactant present with the luminophore during electrochemiluminescence. GAS095.1 7 Preferably, the photodiodes have a planar active surface region for receiving light from the luminophore, and the working electrode has a surface region optically coupled to an active surface region of the photodiode The working electrode is configured such that the optically coupled surface area is greater than 50% of the active surface area of the photodiode. -67- 201211533 GAS095.1 8 Preferably, the LOC device also has a polymerase interlocking reaction (PCR) portion for amplifying the target nucleic acid sequence in the sample. GAS 095.1 9 Preferably, the PCR portion has a heater element for heating the nucleic acid sequence circulating the target and a polymerase, the heater element being configured to be operationally controlled by the C Ο S circuit. GAS095.20 Preferably, the LOC device also has a plurality of sensors coupled to the CMOS circuit for feedback control of the electrodes and the heater elements. The integrated light sensor has the advantage of higher light efficiency than the off-chip sensor design. Integrated light sensors have the advantage of being easier to synchronize with other system events. Integrated light sensors have the advantage of reducing the number of discrete components. Electrochemiluminescence has the advantage of producing effective light at controlled locations in a microfluidic environment. In addition, synchronizing with the sensor is more convenient than a technology system such as fluorescent. This enables more sensitive and specific detection of the target DNA. This LOC device has the advantage of less complex design and manufacturing requirements and therefore results in a simpler, more reliable manufacturing. This LOC device design has the advantage of increasing the coupling between the light emitting region and the light sensor. GAS096.1 This aspect of the invention provides a wafer-on-lab (LOC) device for detecting a target nucleic acid sequence in a sample, the LOC device comprising: having a nucleic acid sequence complementary to the target for forming a probe-target a nucleic acid sequence of the hybrid and a probe of an electrochemiluminescence (ECL) luminophore; and an electrode for generating an excited state of the ECL luminophore, wherein the ECL luminophore emits photons of light in the excited state -68-201211533; Wherein the electrodes are arranged in pairs, one of the electrode pairs causing oxidation or reduction of the luminophore to produce a working electrode of an excited species that emits photons, the working electrode being capable of being emitted by the ECL luminophore The light of the wavelength of the photon penetrates. GAS 096.2 Preferably, each of the probes has a functional portion that is resonated by resonance energy to quench photon emission from the ECL luminophore. φ GAS 096.3 Preferably, the probe is configured such that when the probe forms a probe-target hybrid, the functional moiety for quenching photon emission from the ECL luminophore is further derived from ECL luminophore. GAS 096.4 Preferably, the LOC device also has a CMOS circuit configured to provide electrical pulses to the electrodes. GAS 096.5 Preferably, the electrical pulse has a period of less than 0.69 seconds. GAS096.6 Preferably, the electrical current of the electrical pulse is between 0.1 nanoamperes and 69.0 nanoamperes. GAS 096.7 Preferably, the electrodes have an anode and a cathode, each having a finger configuration, the finger structures being configured to intersect the fingers of the anode with the fingers of the cathode. GAS 096.8 Preferably, the anode and cathode are separated by a dielectric gap of 0.4 microns to 2.0 microns wide. GAS096.9 Preferably, the luminophore is an organometallic complex. GAS 096.1 0 Preferably, the organometallic complex is an organogermanium complex molecule. • 69 201211533 GAS096.11 Preferably, the CMOS circuit has a light sensor for sensing photons emitted by the ECL luminophore. GAS096.1 2 Preferably, the LOC device also has an array of hybridization chambers, wherein each of the hybridization chambers has a pair of electrodes and contains a plurality of probes, and the nucleic acid sequence of the probes in each of the hybridization chambers The nucleic acid sequences in at least one other hybridization chamber in the array are different such that a plurality of target nucleic acid sequences can be detected. GAS 096.1 3 Preferably, the LOC device also has a support substrate, wherein the CMOS circuit is located between the hybridization chamber and the support substrate such that the light sensor is adjacent to the hybridization chambers. GAS096.1 4 Preferably, the light sensor is an array of photodiodes, and the position of the photodiode array is configured such that each of the photodiodes respectively corresponds to one of the hybrid chambers . GAS096.1 5 Preferably, the photodiodes have planar active surface regions for receiving light from the luminophore, each of the active surface regions being in the same plane, and the layers of the electrodes are patterned Patterned to form the conductive material of the separate anode and cathode, the plane of the layer being parallel to the plane of the active surface region of the photodiodes 〇GAS096.1 6 preferably, each of the electrodes is An electrode system causes oxidation or reduction of the luminophore to produce a working electrode of an excited species that emits photons, the working electrode being positioned such that the probes are interposed between the photodiode and the working electrode. GAS 096. 1 7 Preferably, the photodiodes have a planar active surface region for receiving light from the illuminating group of -70-201211533, and the working electrode has an active surface region with the photodiode The surface area of the optical coupling, the working electrode being configured such that a surface area optically coupled to the active surface area is greater than 50% of an active surface area of the photodiode. GAS 096.1 8 Preferably, the LOC device also has a polymerase interlocking reaction (PCR) portion for amplifying the target nucleic acid sequence in the sample. φ GAS 096.1 9 Preferably, the PCR section has a heater element for heating the nucleic acid sequence circulating the target and a polymerase, the heater element being configured to be operationally controlled by the CMOS circuit. GAS096.20 Preferably, the LOC device also has a plurality of sensors coupled to the CMOS circuit for feedback control of the electrodes and the heater elements. The integrated light sensor has the advantage of higher light efficiency than the off-chip sensor design. The integrated light sensor has the advantage of being easier to synchronize with other system events φ. Integrated light sensors have the advantage of reducing the number of discrete components. Electrochemiluminescence has the advantage of producing effective light at controlled locations in a microfluidic environment. In addition, synchronizing with the sensor is more convenient than a technology system such as fluorescent. This enables more sensitive and specific detection of the target DNA. This LOC device has the advantage of less complex design and manufacturing requirements and therefore results in a simpler, more reliable manufacturing. This L0C device design has the advantage of increasing the affinity between the light emitting region and the light sensor. GAS097.1 This aspect of the invention provides a microfluidic device for detecting a target nucleic acid sequence in a sample, the microfluidic device comprising: -71 - 201211533 reagent reservoir for use in detecting the target nucleic acid sequence Adding a reagent to the sample; a probe having a nucleic acid sequence complementary to the target nucleic acid sequence for forming a probe-target hybrid and an electrochemiluminescence (ECL) luminophore; and an excitation state for generating an ECL luminophore An electrode in which the ECL luminophore emits photons of light. GAS 097.2 Preferably, each of the probes has a functional portion that is resonated by resonance energy to quench photon emission from the ECL luminophore. GAS 0 9 7 · 3 Preferably, the probe is configured such that when the probe forms a probe-target hybrid, the functional moiety for quenching photon emission from the ECL luminophore is further From the ECL luminophore. GAS 097.4 Preferably, the microfluidic device also has a CMOS circuit configured to provide electrical pulses to the electrodes. GAS 097.5 Preferably, the electrical pulse has a period of less than 0.69 seconds. GAS097.6 Preferably, the current of the electrical pulse is between 0.1 nanoamperes and 69.0 nanoamperes. GAS 097.7 Preferably, the electrodes have an anode and a cathode, each anode and cathode having a finger configuration configured to intersect each other. GAS097.8 Preferably, the anode and cathode are separated by a dielectric gap of 0.4 microns to 2.0 microns. GAS097.9 preferably the microfluidic device also has a support substrate for supporting the CMOS circuit and defining the reagent reservoir upper cover therein, wherein the electrodes and the probes are interposed between the upper cover and the CMOS Circuit between -72- 201211533. GAS097.10 Preferably, the reagent reservoirs each have an outlet valve for retaining a liquid reagent in the reservoir until a reagent needs to be added to the sample. GAS097.1 1 Preferably, the CMOS circuit has light sensing For sensing photons emitted by the ECL luminophore. GAS097.1 2 Preferably, the microfluidic device also has an array of φ of hybridization chambers, wherein each of the hybridization chambers has a pair of electrodes and contains a plurality of probes, and the nucleic acid sequence of the probes in each of the hybridization chambers The nucleic acid sequence is different from the nucleic acid sequence in at least one other hybridization chamber in the array such that a plurality of target nucleic acid sequences can be detected. GAS 097.1 3 Preferably, the microfluidic device also has a support substrate, wherein the CMOS circuit is located between the hybridization chamber and the support substrate such that the light sensor is adjacent to the hybridization chambers. Preferably, the light sensor is an array of photodiodes that are aligned with the array φ of the hybridization chamber such that each of the hybridization chambers corresponds to one of the photodiodes. GAS097.1 5 Preferably, the photodiodes have planar active surface regions for receiving light from the luminophore, each of the active surface regions being in the same plane, and the layers of the electrodes are patterned Patterned to form the conductive material of the separate anode and cathode, the plane of the layer being parallel to the plane of the active surface region of the photodiodes 〇GAS097.1 6 preferably, each of the electrodes is An electrode system -73-201211533 results in oxidation or reduction of the luminophore to produce a working electrode of an excited species that emits photons, the working electrode being positioned such that the probes are interposed between the photodiode and the work Between the electrodes. GAS097.1 7 Preferably, the photodiodes have a planar active surface region for receiving light from the luminophore, and the working electrode has a surface region optically coupled to an active surface region of the photodiode The working electrode is configured such that the optically coupled surface area is greater than 50% of the active surface area of the photodiode. GAS 097.1 8 Preferably, the microfluidic device also has a polymerase interlocking reaction (PCR) portion for amplifying the target nucleic acid sequence in the sample. GAS 097.1 9 Preferably, the PCR portion has a heater element for heating the nucleic acid sequence circulating the target and a polymerase, the heater element being configured to be operationally controlled by the CMOS circuit. GAS097.20 Preferably, the microfluidic device also has a plurality of sensors coupled to the CMOS circuit for feedback control of the electrodes and the heater elements. The detection and analysis of the subject based on electricity and chemiluminescence eliminates the need for a detection system that requires an excitation source, an excitation optical, and an optical filter element, thereby providing a lighter and less expensive detection system. It is not necessary to reject any excitation light and simplify the detector circuit, making the detection system even cheaper. GAS 098.1 Aspects of the invention provide a microfluidic device for detecting a target molecule in a fluid, the microfluidic device comprising: a reagent reservoir for adding a reagent -74 - 201211533 to the detection of the target molecule to the a fluid; a probe that reacts with the target molecule to form a probe-target hybrid; and an electrochemiluminescence (ECL) luminophore; an electrode for generating an excited state of the ECL luminophore, in which the ECL luminophore emits a photon of light; and a light sensor for sensing photons emitted from the ECL luminophore 〇 φ GAS 098.2 Preferably, each of the probes has resonance energy transfer to quench the luminescent group from the ECL The functional part of photon emission. GAS 098.3 Preferably, the probe is configured such that when the probe forms a probe-target complex, the functional moiety for quenching photon emission from the ECL luminophore is further derived from the ECL Luminous group. GAS 098.4 Preferably, the microfluidic device also has a CMOS circuit configured to provide electrical pulses to the electrodes. GAS098.5 Preferably, the electrical pulse has a period of φ of less than 0.69 seconds. GAS098.6 Preferably, the electrical current of the electrical pulse is between 0.1 nanoamperes and 69.0 nanoamperes. GAS 098.7 Preferably, the electrodes have an anode and a cathode, each having a finger configuration, the finger structures being configured to intersect the fingers of the anode with the fingers of the cathode. GAS098.8 Preferably, the anode and cathode are separated by a dielectric gap of 0.4 microns to 2.0 microns. GAS09 8.9 Preferably, the microfluidic device also has a support substrate for supporting the 201211533 CMOS circuit and defining the reagent reservoir upper cover therein, wherein the electrodes and the probes are interposed between the upper cover and the CMOS Between circuits. GAS 098.1 0 Preferably, the reagent reservoirs each have an outlet valve for retaining a liquid reagent in the reservoir until a reagent needs to be added to the fluid 〇GAS 09 8.il. Preferably, the CMOS circuit has The light sensor is such that the light sensor is in close proximity to the hybrid chambers. GAS098.1 2 Preferably, the microfluidic device also has an array of hybridization chambers, wherein each of the hybridization chambers has a pair of electrodes and comprises a plurality of probes, wherein the flow system biological sample and the labeled labeled nucleic acids a sequence, each of which has a nucleic acid sequence complementary to each of the targets 〇 GAS 098.1 3 Preferably, the microfluidic device also has a support substrate, wherein the CMOS circuit is located in the hybrid chamber and The substrates are supported to position the light sensor adjacent to the hybrid chambers. GAS 098.1 4 Preferably, the light sensor is an array of photodiodes locating the hybridization chamber such that each of the hybridization chambers corresponds to one of the photodiodes. GAS 098.1 5 Preferably, the photodiodes have planar active surface regions for receiving light from the luminophore, each of the active surface regions being in the same plane, and the layers of the electrodes being patterned ( Patterned to form a conductive material of the separate anode and cathode, the plane of the layer being parallel to the plane of the active surface region of the photodiode - 76 - 201211533 GAS098.16 Preferably, each of the electrodes is centered One of the electrodes causes oxidation or reduction of the luminophore to produce a working electrode of an excited species that emits photons, the working electrode being positioned such that the probes are interposed between the photodiode and the working electrode . GAS098.1 7 Preferably, the photodiodes have a planar active surface region for receiving light from the luminophore, and the working electrode has a light coupling with an active surface region of the photodiode The surface region, the working electrode is configured such that the optically coupled surface area is greater than 50% of the active surface area of the photodiode. GAS098.1 8 Preferably, the microfluidic device also has a polymerase chain reaction (PCR) portion for amplifying the target nucleic acid sequence in the sample. GAS 098.1 9 Preferably, the PCR portion is useful The target nucleic acid sequence is heated and heated to a heater element of the polymerase, the heater element φ being configured to be operationally controlled by the CMOS circuit. GAS098.20 Preferably, the microfluidic device also has a plurality of sensors coupled to the CMOS circuit for feedback control of the electrodes and the heater elements. The integrated image sensor eliminates the need for expensive external imaging systems, providing a comprehensive solution that is both mass-produced and inexpensive, with low system component counts for lightweight, highly mobile systems. The integrated image sensor has the benefit of increased read sensitivity due to large angle light collection and eliminates the need to use optical components in the string of light collecting elements. -77- 201211533 This electrochemiluminescence-based detection analysis eliminates the need for detection systems that require excitation light sources, excitation optics, and optical filter elements to provide a lighter and less expensive inspection system. It is not necessary to reject any excitation light and simplify the detector circuit, making the detection system even cheaper. GAS099.1 This aspect of the invention provides a microfluidic device comprising: a support substrate; a microsystem technology (MST) layer provided with a hybrid portion having an array of electrochemiluminescence (ECL) probes for use with Hybridization of a target nucleic acid sequence in a fluid, and an electrode pair for receiving an electrical pulse, the ECL probes being configured to hybridize upon activation with one of the nucleic acid targets and upon activation by one of the electrodes a photon of light; and a light sensor for detecting photons of light from an already hybridized ECL probe. GAS 099.2 Preferably, the probes each have an ECL luminophore that emits photons when in an excited state and a functional moiety for photon emission from the ECL luminophore by resonance energy transfer quenching. GAS 099.3 Preferably, the probes are configured such that when the probe forms a probe-target hybrid, the functional moiety for quenching photon emission from the ECL luminophore is further derived The ECL luminophore. GAS 099.4 Preferably, the microfluidic device also has a CMOS circuit configured to provide electrical pulses to the electrodes. GAS099.5 Preferably, the electrical pulse has a duration of less than 0.69 seconds 201211533 GAS099.6 Preferably, the electrical current of the electrical pulse is between 0.1 nanoamperes and 69.0 nanoamperes. GAS 099.7 Preferably, the electrodes have an anode and a cathode, each having a finger configuration, the finger structures being configured to intersect the fingers of the anode with the fingers of the cathode. GAS 099.8 Preferably, the anode and cathode are separated by a dielectric gap of between 0.4 microns and 2.0 microns. GAS099.9 Preferably, the microfluidic device also has a support substrate for supporting the CMOS circuit and defining the reagent reservoir upper cover therein, wherein the electrodes and the probes are interposed between the upper cover and the CMOS Between circuits. Preferably, the upper cap has a reagent reservoir for adding a reagent to the sample prior to detecting the target nucleic acid sequence, each reagent reservoir having an outlet valve for retaining a liquid reagent within the reservoir Until the reagent needs to be added to the sample. GAS099.il Preferably, the reagent reservoirs each have an outlet valve for retaining a liquid reagent in the reservoir until a reagent needs to be added to the sample 〇GAS099.12. Preferably, the microfluidic device also has a hybridization chamber. An array wherein each of the hybridization chambers has a pair of electrodes and a plurality of probes, and the nucleic acid sequence of the probes in each of the hybridization chambers is different from the nucleic acid sequence in at least one other hybridization chamber in the array, In order to allow a plurality of target nucleic acid sequences to be detected. Preferably, the microfluidic device also has a support substrate -79-201211533, wherein the CMOS circuit is located between the hybridization chamber and the support substrate such that the light sensor is adjacent to the hybrid chambers. . GAS 099.14 Preferably, the light sensor is an array of photodiodes locating the hybridization chamber such that each of the hybridization chambers corresponds to one of the photodiodes. GAS099.1 5 Preferably, the photodiodes have a planar active surface region for receiving light from the luminophore, each of the active surface regions being in the same plane, and the electrodes are patterned by a layer Patterned to form the conductive material of the separate anode and cathode, the plane of the layer being parallel to the plane of the active surface region of the photodiodes. GAS099.1 6 Preferably, each of the electrodes is centered One of the electrodes causes oxidation or reduction of the luminophore to produce a working electrode of an excited species that emits photons, the working electrode being positioned such that the probes are interposed between the photodiode and the working electrode . GAS099.1 7 Preferably, the photodiodes have a planar active surface region for receiving light from the luminophore, and the working electrode has a surface region optically coupled to an active surface region of the photodiode The working electrode is configured such that the optically coupled surface area is greater than 50% of the active surface area of the photodiode. GAS 099.1 8 Preferably, the microfluidic device also has a polymerase interlocking reaction (PCR) portion for amplifying the target nucleic acid sequence in the sample. GAS099.19 Preferably, the PCR portion has a heater element for heating the circulating nucleic acid sequence 201211533 with a polymerase, the heater element being configured to be operationally controlled by the CMOS circuit. GAS099.20 Preferably, the microfluidic device also has a plurality of sensors connected to the CMOS circuit for feedback control of the electrodes and the heater element. The sample inlet can introduce the sample into the microfluidic test. A module that delivers a small amount of sample to a designated portion of the microfluidic device with high volumetric efficiency. The probe hybrid provides analysis of the target via hybridization. The electrochemiluminescence-based detection analysis eliminates the need for a detection system that requires an excitation source, excitation optics, and optical over-the-loop components to provide a lighter and less expensive inspection system. It is not necessary to reject any excitation light and simplify the detector circuit, making the detection system even cheaper. GAS100.1 This aspect of the invention provides a microfluidic device for detecting a target nucleic acid sequence in a sample, the microfluidic device comprising: a sample into the sample for accepting the sample; indeed complementary to the target nucleic acid sequence a probe for forming a nucleic acid sequence of a probe-target hybrid and an electrochemiluminescence (ECL) luminophore; and an electrode for generating an excited state of the ECL luminophore, wherein the ECL luminophore emits light in the excited state Photon; wherein the sample inlet attracts the sample along the fluid flow path leading to the probes by capillary action. GAS 100.2 Preferably, each of the probes has a functional portion that is resonated by resonance energy to quench photon emission from the ECL luminophore. GAS 100.3 Preferably, the probe is configured such that when the probe -81 - 201211533 forms a probe-target hybrid, the functional portion for quenching photon emission from the ECL luminophore is further From the ECL luminophore. GAS 100.4 Preferably, the microfluidic device also has a CMOS circuit configured to provide electrical pulses to the electrodes. GAS 1 00.5 Preferably, the electrical pulse has a period of less than 〇 · 6.9 seconds. Preferably, the electrical current of the electrical pulse is between 0.1 nanoamperes and 69.0 nanoamperes. GAS 1 00.7 Preferably, the electrodes have an anode and a cathode, each having a finger configuration, the finger structures being configured to intersect the fingers of the anode with the fingers of the cathode. GAS 100.8 Preferably, the anode and cathode are separated by a dielectric gap of 0.4 microns to 2.0 microns. GAS100.9 Preferably, the microfluidic device also has a support substrate for supporting the CMOS circuit and defining the reagent reservoir upper cover therein, wherein the electrodes and the probes are interposed between the upper cover and the CMOS Between circuits. GAS 100.10 Preferably, the upper cap has a reagent reservoir for adding a reagent to the sample prior to detecting the target nucleic acid sequence, each of the reagent reservoirs having an outlet valve for retaining a liquid reagent in the reservoir Add reagents to the sample until needed. Preferably, the reagent reservoirs each have an outlet valve for retaining a liquid reagent in the reservoir until a reagent needs to be added to the sample - 82 - 201211533 GAS 100.12 Preferably, the microfluidic device is also An array having a hybridization chamber, wherein each of the hybridization chambers has a pair of electrodes and a plurality of probes, and the nucleic acid sequence of the probes in each of the hybridization chambers and the nucleic acid in at least one other hybridization chamber in the array The sequences are different such that a plurality of target nucleic acid sequences can be detected. GAS100.13 Preferably, the microfluidic device also has a light sensor and a support substrate for sensing photons emitted from the ECL luminophore, wherein the CMOS circuit is located between the hybridization chamber and the support substrate So that the light sensor is adjacent to the hybrid chambers. GAS 100.14 Preferably, the light sensor is an array of photodiodes, the photodiode array being positioned such that each of the photodiodes corresponds to one of the hybridization chambers. GAS 100.15 Preferably, the photodiodes have planar active surface regions for receiving light from the luminophore, each of the active surface regions being in the same plane, and the layers of the electrodes are patterned ^ ( Patterned to form a conductive material of the separate anode and cathode, the plane of the layer being parallel to the plane of the active surface region of the photodiodes. · GAS 1 00.1 6 Preferably, each of the pairs of electrodes One of the electrodes causes oxidation or reduction of the luminophore to produce a working electrode of an excited species that emits photons, the working electrode being positioned such that the probes are interposed between the photodiode and the working electrode between. GAS100.17 Preferably, the photodiodes have a planar active surface area for receiving light from the luminophore, and the working electrode has an active surface area light with the photodiode -83-201211533 The coupled surface region is configured such that the optically coupled surface region is greater than 50% of the active surface region of the photodiode. GAS 100.18 Preferably, the microfluidic device also has a polymerase interlocking reaction (PCR) portion for amplifying the target nucleic acid sequence in the sample. GAS 100.19 Preferably, the PCR portion has a heater element for heating the nucleic acid sequence circulating the target and a polymerase, the heater element being configured to be operationally controlled by the CMOS circuit. GAS 1 00.20 Preferably, the microfluidic device also has a plurality of sensors coupled to the CMOS circuit for feedback control of the electrodes and the heater elements. The probe hybridization provides analysis of the target via hybridization. The integrated image sensor eliminates the need for expensive external imaging systems, providing a comprehensive solution that is both mass-produced and inexpensive, with low system component counts for lightweight, highly mobile systems. The integrated image sensor has the benefit of increased read sensitivity due to large angle light collection and eliminates the need to use optical components in the light collecting element string. The electrochemiluminescence-based detection analysis eliminates the need for detection systems that require excitation light sources, excitation optics, and optical filter elements to provide a lighter and less expensive inspection system. It is not necessary to reject any excitation light and simplify the detector circuit, making the detection system even cheaper. GAS 1 01 . 1 This aspect of the invention provides a microfluidic device for detecting a target nucleic acid sequence in a sample, the microfluidic device comprising: -84 - 201211533 a support substrate; covering the support substrate for processing the sample a microsystem technology (MST) layer, the MST layer being provided with an array of hybridization chambers, each hybridization chamber containing an electrochemiluminescence (ECL) probe for hybridization with the target nucleic acid sequence, and an electrode pair for receiving an electrical pulse The ECL probes are configured to emit photons of light upon hybridization with one of the nucleic acid targets and upon activation by one of the electrodes; an array of temperature sensors arranged at a location Having at least one of the temperature sensors corresponding to each of the hybridization chambers; and a heater for heating each of the hybridization chambers; such that outputs from the temperature sensors are used to feedback control the heaters. GAS 101.2 Preferably, the microfluidic device also has a CMOS circuit between the support substrate and the MST layer, the CMOS circuit having a light sensor. GAS 101.3 Preferably, the light sensor is an array of photodiodes respectively registered with each of the hybrid chambers. GAS 101.4 Preferably, the CMOS circuit has digital memory for storing data relating to the processing of the sample, the data including the probe details and the location of each of the probes in the array. GAS 10 1.5 Preferably, the microfluidic device also has a reagent reservoir that integrally contains all reagents for processing the sample.

GAS 101.6 Preferably, the hybridization chamber has a heater controlled by the CMOS circuit, respectively, for maintaining the hybridization temperature of the probe and the target nucleic acid sequence -85 - 201211533. GAS 101.7 Preferably, the light sensor is less than 1,600 microns from the corresponding hybridization chamber. GAS 1 01 .8 Preferably, the hybridization chamber has a volume of less than 900,000 cubic microns. GAS 1 01.9 Preferably, the hybridization chamber has a volume of less than 200,000 cubic microns.

GAS 1 01.1 0 Preferably, the hybridization chamber has a volume of less than 40,000 cubic microns. GAS 101.il Preferably, the hybridization chamber has a volume of less than 9,000 cubic microns. GAS 101.12 Preferably, the CMOS circuit derives a single result from a photodiode corresponding to a hybridization chamber containing the same probe. GAS 101.13 Preferably, the CMOS circuit is configured to provide an excitation pulse to the pair of electrodes, the period of the excitation pulse being less than 0.69 seconds.

Preferably, the light sensor is an array of photodiodes, the photodiode array being positioned such that each of the photodiodes corresponds to one of the hybrid chambers. Preferably, the photodiodes have planar active surface regions for receiving light from the luminophore, each of the active surface regions being in the same plane, and the layers of the electrodes being patterned ( Patterned to form a conductive material of the separate anode and cathode, the plane of the layer being parallel to the plane of the active surface region of the photodiodes - 86 - 201211533 GAS 10 1.1 6 Preferably, each of the pairs of electrodes One of the electrodes causes oxidation or reduction of the luminophore to produce a working electrode of an excited species that emits photons, the working electrode being positioned such that the probes are interposed between the photodiode and the working electrode between. Preferably, the photodiodes have a planar active surface region for receiving light from the luminophore, and the working electrode has a surface region optically coupled to an active surface region of the photodiode, The working electrode is configured such that the optically coupled surface area is greater than 50% of the active surface area of the photodiode. GAS 10 1.1 8 Preferably, the microfluidic device also has a polymerase interlocking reaction (PC R) portion for amplifying the target nucleic acid sequence in the sample. GAS 101.19 Preferably, the PCR portion has a heater element for heating the nucleic acid sequence circulating the target and a polymerase, the heater element being configured to be operationally controlled by the CMOS circuit. GAS 101.20 Preferably, the microfluidic device also has a plurality of sensors coupled to the CMOS circuit for feedback control of the electrodes and the heater elements. The probe hybridization provides analysis of the target via hybridization. This temperature feedback control ensures control of the temperature within the hybrid chamber to achieve optimal hybridization temperature and subsequent optimal detection temperature. The electrochemiluminescence-based detection analysis eliminates the need for detection systems that require excitation light sources, excitation optics, and optical filter elements to provide a lighter and less expensive inspection system. There is no need to reject any excitation light. -87-201211533 This detector circuit makes the detection system even cheaper. GAS 1 02.1 This aspect of the invention provides a microfluidic device for detecting a target nucleic acid sequence in a sample, the microfluidic device comprising: an array of hybridization chambers, each hybridization chamber containing an electrochemical for hybridization to the target nucleic acid sequence An illuminating (ECL) probe, and an electrode pair for receiving an electrical pulse, the ECL probes being configured to hybridize upon activation with one of the nucleic acid targets and upon activation by one of the electrodes Photon of light; wherein each of the hybridization chambers has a wall portion that is transparent to light emitted by the ECL probe. GAS 102.2 Preferably, the microfluidic device also has a light sensor for detecting light emitted by the ECL probe, wherein the wall portion is between the ECL probe and the light sensor. Preferably, the microfluidic device also has the light sensor and the support substrate of the hybrid chamber array, wherein the light sensor is located between the hybridization chamber and the support substrate, and the wall portion is There is a layer of cerium oxide. The GAS 102·4 preferably 'the probes each have an ECL luminophore that emits photons when in an excited state and a functional moiety for photon emission from the ECL luminophore by resonance energy transfer quenching. GAS 102.5 Preferably, the probes are configured such that when the probe forms a probe-target hybrid, the functional moiety for quenching photon emission from the ECL luminophore is further derived from ECL luminophore. GAS 1 02.6 Preferably the microfluidic device also has a CMOS circuit configured to provide electrical pulses to the electrodes. - 88 - 201211533 GAS 102.7 Preferably, the electrical pulse has a period of less than 0.69 seconds. Preferably, the electrical current of the electrical pulse is between 0.1 nanoamperes and 69.0 nanoamperes. GAS 102.9 Preferably, the electrodes have an anode and a cathode, each having a finger configuration, the finger structures being configured to intersect the fingers of the anode with the fingers of the cathode. GAS 102.10 Preferably, the anode and cathode are separated by a dielectric gap of from 4 micrometers to 2.0 micrometers wide. Preferably, the microfluidic device also has an upper cover, the upper cover being provided with a reagent reservoir for adding to the sample prior to detecting the sequence of the target, wherein the electrodes and the probes are interposed Between the upper cover and the CMOS circuit. Preferably, the reagent reservoirs each have an outlet valve for retaining a liquid reagent in the reservoir until a reagent needs to be added to the sample 〇GAS 102.13. Preferably, the transparent wall portion has a thickness of less than one. 600 micron" GAS 102.14 Preferably, the light sensor is an array of photodiodes that are positioned in registration with the hybridization chamber such that each of the hybridization chambers corresponds to one of the photodiodes. Preferably, the photodiodes have planar active surface regions for receiving light from the luminophore, each of the active surface regions being in the same plane, and the layers of the electrodes are patterned 89 - 201211533 (patterned) to form the conductive material of the separate anode and cathode, the plane of the layer being parallel to the plane of the active surface region of the photodiode 〇GAS102.16 Preferably, each of the electrodes is centered One of the electrodes causes oxidation or reduction of the luminophore to produce a working electrode of an excited species that emits photons, the working electrode being positioned such that the probes are interposed between the photodiode and the working electrode . GAS 102.17 Preferably, the photodiodes have a planar active surface region for receiving light from the luminophore, and the working electrode has a surface region optically coupled to an active surface region of the photodiode, The working electrode is configured such that the optically coupled surface area is greater than 50% of the active surface area of the photodiode. GAS 102.18 Preferably, the microfluidic device also has a polymerase interlocking reaction (PCR) portion for amplifying the target nucleic acid sequence in the sample. GAS 102.19 Preferably, the PCR portion has a heater element for heating the nucleic acid sequence circulating the target and a polymerase, the heater element being configured to be operationally controlled by the CMOS circuit. GAS 1 02.20 Preferably, the microfluidic device also has a plurality of sensors coupled to the CMOS circuit for feedback control of the electrodes and the heater elements. The probe hybridization provides analysis of the target via hybridization. The light-transmissive hybridization chamber provides for delivery of an electrochemical luminescence signal for detecting hybridization of the target with the probe. 201211533 This electrochemiluminescence-based detection analysis eliminates the need for detection systems that require excitation light sources, excitation optics, and optical filter elements to provide a lighter and less expensive inspection system. It is not necessary to reject any excitation light and simplify the detector circuit, making the detection system even cheaper. GAS 1 03.1 This aspect of the invention provides a microfluidic device for detecting a target nucleic acid sequence in a sample, the microfluidic device comprising:

An array of hybridization chambers - each of the hybridization chambers comprising an electrode pair for receiving an electrical pulse and an electrochemiluminescence (ECL) probe for hybridizing to the target nucleic acid sequence, the ECL probes being configured such that A photon that hybridizes to one of the nucleic acid targets and emits light when excited by a current between the electrodes; wherein the probe mass in each of the hybridization chambers is less than 270 picograms. GAS 103.2 Preferably, the probe mass in each of the hybridization chambers is less than 60 picograms. GAS 103.3 Preferably, the probe mass in each of the hybridization chambers is less than 12 picograms. GAS 103.4 Preferably, the probe mass in each of the hybridization chambers is less than 2.7 picograms. GAS 103.5 Preferably, each of the hybrid chambers comprises one of the pair of electrodes.

GAS103. Preferably, the hybridization chambers each have a wall portion that is permeable to light emitted by the ECL probe. GAS 103. Preferably, the microfluidic device also has a light sensor for detecting light emitted by the ECL probe. -91 - 201211533 GAS 103. Preferably, the probes each have an ECL luminophore that emits photons when in an excited state and a functional moiety for photon emission from the ECL luminophore by resonance energy transfer quenching. GAS 103. Preferably, the probes are configured such that when the probe forms a probe-target hybrid, the functional moiety for quenching photon emission from the ECL luminophore is further derived from the ECL Luminous group. GAS103. Preferably, the microfluidic device also has a CMOS circuit configured to provide electrical pulses to the electrodes. GAS103. Il preferably, the electrical pulse has a magnitude less than zero. During the 69 seconds period. GAS103. 12 Preferably, the current of the electrical pulse is between 0. 1 nai to 69. 0 nai pei. GAS 103.  Preferably, the pair of electrodes have an anode and a cathode, each having a finger configuration, the finger structures being configured to intersect the fingers of the anode with the fingers of the cathode. GAS103. 14 Preferably, the anode and cathode are separated by 0. 4 microns to 2. 0 micron wide dielectric gap. GAS 103.  Preferably, the microfluidic device also has an upper cover, the upper cover being provided with a reagent reservoir for adding to the sample prior to detecting the sequence of the target, wherein the electrodes and the probes are interposed between the upper cover and Between these CMOS circuits. GAS103. Preferably, the reagent reservoirs each have an outlet valve for retaining the liquid reagent in the reservoir until the reagent needs to be added to the sample -92 - 201211533 GAS 1 03. Preferably, the light sensor is an array of photodiodes locating the hybridization chamber such that each of the hybridization chambers corresponds to one of the photodiodes. GAS103. Preferably, the microfluidic device also has a polymerase interlocking reaction (PCR) portion for amplifying the target nucleic acid sequence in the sample. GAS 103. Preferably, the PCR portion has a heater element for heating the cycle φ the nucleic acid sequence of the target and the polymerase. The heater element is configured to be operationally controlled by the CMOS circuit. GAS 1 03. Preferably, the microfluidic device also has a plurality of sensors coupled to the CMOS circuit for feedback control of the electrodes and the heater elements. This low probe volume represents a low probe cost, which in turn allows for this inexpensive detection system. The electrochemiluminescence-based detection analysis eliminates the need for a detection system that illuminates the source, excitation optics, and optical filter elements, thereby providing a lighter and less expensive inspection system. It is not necessary to reject any excitation light and simplify the detector circuit, making the detection system even cheaper. GAS 105. 1 This aspect of the invention provides a microfluidic device for amplifying and detecting a target nucleic acid sequence in a sample, the microfluidic device comprising: a polymerase chain reaction (PC R) portion for amplifying the a nucleic acid sequence; and a hybridization portion having an array of electrochemiluminescence (ECL) probes for hybridizing with the target nucleic acid sequences to form a probe-target hybrid, and a plurality of -93-201211533 for receiving electricity Pulses are used to cause the probe-target hybrids to emit light photon electrodes. GAS105. Preferably, the probes each have an ECL luminophore that emits photons when in an excited state and a functional moiety for photon emission from the ECL luminophore by resonance energy transfer quenching. GAS105. Preferably, the probes are configured such that when the probe forms a probe-target hybrid, the functional moiety for quenching photon emission from the ECL luminophore is further derived from the ECL Luminous group. GAS 105. Preferably, the microfluidic device also has a CMOS circuit configured to provide electrical pulses to the electrodes. GAS105. 5 Preferably, the electrical pulse has a value less than 0. During the 69 seconds period. GAS105. 6 Preferably, the current of the electrical pulse is between 0. 1 nai to 69. 0 nai pei. GAS105. Preferably, the electrodes have an anode and a cathode, each having a finger configuration, the finger structures being configured to intersect the fingers of the anode with the fingers of the cathode. GAS105. 8 Preferably, the anode and the cathode are separated by 0. 4 microns to 2. 0 micron wide dielectric gap. GAS105. Preferably, the microfluidic device also has an upper cover, the upper cover is provided with a reagent reservoir for adding to the sample before detecting the target sequence, wherein the electrodes and the probes are interposed between the upper cover and the probe Between the C Μ Ο S circuits. GAS105. Preferably, the reagent reservoirs each have an outlet valve for -94-201211533 for retaining liquid reagents in the reservoir until reagents need to be added to 〇 GAS 105. Il. Preferably, the microfluidic device also has an array of hybridization chambers for the ECL probes and a pair of electrodes. GAS105. Preferably, the hybridization chambers each have a wall portion that is transparent to the light emitted by the probe. GAS 105. Preferably, the microfluidic device also has a light sensor with φ light emitted by the ECL probe, wherein the wall portion is between the ECL probe and the light sensor. GAS 105. Preferably, the microfluidic device also has a support substrate for the array of the hybridization chamber, wherein the light sensor is between the hybridization chamber and the support substrate, and the wall portion is doped with a dioxide layer. GAS105. Preferably, the light sensor is positioned and aligned with the array of photodiodes such that each of the hybrid chambers is respectively associated with one of the φ photodiodes. GAS 1 05. Preferably, the array of the photodiodes is at a distance of less than 1,6 microns. GAS105. Preferably, the photodiodes have planar active surface regions with light from the luminophores, each of the main regions being in the same plane, and the layers of the electrodes are G (patterned) to form the separation The plane where the conductive material of the anode and the cathode is located is equal to the active surface area of the photodiode, and the sample is included in the chamber containing the ECL at the detection system and is located at the junction of the wafer. Surface B case, the level is parallel -95- 201211533 GAS 105. Preferably, one of the electrode pairs causes oxidation or reduction of the luminophore to produce a working electrode of an excited species that emits photons, the working electrode being configured such that the probes are interposed The photodiode is between the working electrode and the working electrode. GAS 105. Preferably, the PCR portion has a microchannel extending between the PCR inlet and the PCR outlet, the microchannel being configured to attract the sample from the PCR inlet to the PCR outlet by capillary action. GAS 1 05. Preferably, the microchannel has a plurality of elongate heaters, each of the plurality of elongate heaters being independently operable. The PCR section is amplified by the target to provide the sensitivity required for the target detection. The probe hybridization provides analysis of the target via hybridization. The integrated PCR and probe hybridization section substantially reduces the possibility of introducing contamination during the assay, simplifies the analysis phase, and provides a lightweight and inexpensive single-device analysis solution. The electrochemiluminescence-based detection analysis eliminates the need for detection systems that require excitation light sources, excitation optics, and optical filter elements to provide a lighter and less expensive inspection system. It is not necessary to reject any excitation light and simplify the detector circuit, making the detection system even cheaper. GAS 1 06. 1 Aspects of the invention provide a microfluidic device for detecting a target molecule in a fluid, the microfluidic device comprising: an array of chambers, each chamber containing a complex for reacting with the target molecule to form a probe-target complex An electrochemiluminescence (ECL) probe, and an electrode located in each chamber for receiving electrical pulses, the probe-target composite system being configured to emit photons of light when excited by current between the electrodes; -96- 201211533 A flow path containing the fluids of the targets; wherein each of the chambers has a chamber inlet for fluid communication between the sample flow path and a probe in the chamber, the chamber inlet being configured as a diffusion barrier to prevent The probe-target complexes diffuse between the chambers resulting in erroneous detection results. GAS106. 2 Preferably, the chamber inlet defines a tortuous flow path. GAS106. Preferably, the meandering flow path has a curved configuration. φ GAS106. Preferably, the flow system biological sample and the labeled nucleic acid sequences each have a nucleic acid sequence complementary to one of the targets and the chambers are used to hybridize the probes to form Probe-target hybridization chamber. GAS106. Preferably, the hybridization chambers each have a wall portion that is transparent to the light emitted by the probe. GAS106. Preferably, the microfluidic device also has a light sensor for detecting light emitted by the probe, wherein the wall portion is between the φ probe and the light sensor. GAS106. Preferably, the microfluidic device also has a support substrate of the light sensor and the array of hybridization chambers, wherein the light sensor is located between the hybridization chamber and the support substrate, and the wall portion has two Layer of yttrium oxide. GAS106. Preferably, the probes each have an ECL luminophore that emits photons when in an excited state and a functional moiety for photon emission from the ECL luminophore by resonance energy transfer quenching. GAS 106·9 preferably 'the probes are configured such that when the probe-97-201211533 needle forms a probe-target hybrid, the functionality for quenching photon emission from the ECL luminophore Part of the line is further derived from the ECL luminophore. GAS 106. Preferably, the microfluidic device also has a CMOS circuit configured to provide electrical pulses to the electrodes. GAS106. Il preferably, the electrical pulse has a value less than 〇.  6 9 seconds period. GAS106. 12 Preferably, the current of the electrical pulse is between 0. 1 nai to 69. 0 nai pei. GAS106. Preferably, the electrodes have an anode and a cathode, each having a finger configuration, the finger structures being configured to intersect the fingers of the anode with the fingers of the cathode. GAS 106. 14 Preferably, the anode and cathode are separated by 0. Dielectric gap of 4 microns to 2 microns wide. GAS 106. Preferably, the microfluidic device also has an upper cover, the upper cover being provided with a reagent reservoir for adding to the sample prior to detecting the sequence of the target, wherein the electrodes and the probes are interposed between the upper cover and Between the C Μ Ο S circuits. GAS106. 16 Preferably, the reagent reservoirs each have an outlet valve for retaining a liquid reagent in the reservoir until a reagent needs to be added to the sample 〇 GAS 106. Preferably, the layer has a thickness of less than 1,600 microns. GAS 106. Preferably, the light sensor is positioned as an array of photodiodes in registration with the hybridization chamber such that each of the hybridization chambers corresponds to one of the photodiodes. 98- 201211533 GAS106. Preferably, the photodiodes have planar active surface regions for receiving light from the luminophore, each of the active surface regions being in the same plane, and the layers of the electrodes are patterned. To form the conductive material of the separated anode and cathode, the plane of the layer is parallel to the plane of the active surface region of the photodiodes 〇GAS 1 06. Preferably, one of the electrode pairs φ causes oxidation or reduction of the luminophore to produce a working electrode of an excited species that emits photons, the working electrode being positioned such that the probes are interposed Between the photodiode and the working electrode. The probe hybridization provides analysis of the target via hybridization. The diffusion barrier substantially eliminates the reflux of the probe prior to and after hybridization to prevent signal loss and provide a high degree of detection sensitivity. The electrochemiluminescence-based detection analysis eliminates the need for a detection system that requires an excitation source, excitation optics, and optical filter components to provide a lighter and less expensive detection system. It is not necessary to reject any excitation light and simplify the detector circuit, making the detection system even cheaper. GAS 108.  1 Aspects of the invention provide a microfluidic device for detecting a target molecule in a fluid, the microfluidic device comprising: an electrochemiluminescence (ECL) probe for reacting with the target molecule to form a probe - An array of chambers of the target complex; electrodes located in each of the chambers for receiving electrical pulses, the probe-target composite system being configured to emit photons of light when excited by current between the electrodes; -99- 201211533 A light sensor for detecting light emitted by the probes; wherein the light sensor is at a distance of less than 1,600 microns from the probes. GAS108. Preferably, the probe mass in each of the chambers is less than 270 picograms. GAS108. Preferably, the probe mass in each of the chambers is less than 60 picograms. GAS108. Preferably, the probe mass in each of the chambers is less than 12 picograms. GAS108. Preferably, the microfluidic device also has: a support substrate: a CMOS circuit for supplying electrical pulses to the electrodes; and a flow path containing the target fluid; wherein the CMOS is interposed between the chambers and the The substrates are supported, and the flow path attracts the fluid to each of the chambers by capillary action. GAS 108. Preferably, the flow system biological sample and the target nucleic acid sequence, each of the probes has a nucleic acid sequence complementary to the target and the chambers are used to hybridize the probes to form a probe-target hybrid Hybrid room. GAS108. Preferably, the hybridization chambers each have a wall portion that is transparent to the light emitted by the probe. GAS108. Preferably, the CMOS circuit is provided with the light sensor, wherein the wall portion is located between the probes and the light sensor. GAS 108. Preferably, the wall portion is doped with a layer of cerium oxide. GAS108. Preferably, the probes each have an ECL luminophore that emits photons when excited by a current between the electrodes -100 - 201211533 and a photon emission from the ECL luminophore by quenching by resonance energy transfer. The functional part is 〇GAS108. Il. Preferably, the probes are configured such that when the probe forms a probe-target hybrid, the functional moiety for quenching photon emission from the ECL luminophore is further derived from the ECL Luminous group. GAS108. Preferably, the CMOS circuit is configured to provide electrical pulses to the electrodes. GAS 108. 13 Preferably, the electrical pulse has a value less than 0. During the 69 seconds period. GAS 10 8. 14 Preferably, the current of the electrical pulse is between 0. 1 nai to 69. 0 nai pei. GAS108. Preferably, the electrodes have an anode and a cathode, each having a finger configuration, the finger structures being configured to intersect the fingers of the anode with the fingers of the cathode. GAS 108. 16 Preferably, the anode and the cathode are separated by 0. Dielectric gap of 4 microns to 2 microns wide. GAS108. Preferably, the transparent wall portion has a thickness of less than 1,600 microns. GAS108. Preferably, the light sensor is an array of photodiodes locating the hybridization chamber such that each of the hybridization chambers corresponds to one of the photodiodes. GAS108. Preferably, the photodiodes have planar active surface regions for receiving light from the luminophore, each of the active surfaces 101 - 201211533 regions being in the same plane 'and the electrodes are patterned Patterned to form the separate anode and cathode conductive material, the plane of the layer is parallel to the plane of the active surface area of the photodiode GAS108. Preferably, one of the electrode pairs causes oxidation or reduction of the luminophore to produce a working electrode of an excited species that emits photons, the working electrode being configured such that the probes are interposed The photodiode is between the working electrode and the working electrode. This non-imaging optics provides a comprehensive solution for mass production and inexpensive, low system components, and provides a lightweight, highly mobile system. This non-imaging optics provides the benefit of increased read sensitivity due to large angle light collection and eliminates the need to use optical components in the string of light collecting elements. The electrochemiluminescence-based detection analysis eliminates the need for detection systems that require excitation light sources, excitation optics, and optical filter elements to provide a lighter and less expensive inspection system. It is not necessary to reject any excitation light and simplify the detector circuit, making the detection system even cheaper. GAS 1 09. 1 Aspects of the invention provide a microfluidic device for detecting a target molecule in a fluid, the microfluidic device comprising: a hybridization chamber comprising an electrode and a probe for hybridizing with a target molecule to form a probe-target The composite and the current between the electrodes are returned to generate an electrochemiluminescence (ECL) signal; and the photodiode has an active region and an optical axis extending perpendicular to the active region and passing through the hybridization chamber; wherein the hybridization chamber a surface having a bottom portion 8-102-201211533 parallel to the active region of the photodiode, the bottom surface having a center of mass and the active region being included in a pyramid having a center of mass at a vertex thereof, and a vertex angle The system is less than 1 7 4 . . GAS 109. 2 Preferably, the apex angle is less than 29 degrees. GAS 109. 3 Preferably, the apex angle is less than 4. 8 degrees. GAS109. 4 Preferably, the apex angle is less than 0. 8 degrees. GAS109. Preferably, the microfluidic device also has: a support substrate; a CMOS circuit for supplying electrical pulses to the electrodes; an array of the hybridization chambers; and a flow path of a fluid containing the target molecules; wherein the CMOS A circuit is interposed between the chambers and the support substrate, and the flow path attracts the fluid to each of the chambers by capillary action. GAS109. Preferably, the flow system biological sample and the target nucleic acid sequence, each of the probes has a nucleic acid sequence complementary to the target and the chambers are used to hybridize the probes to form a probe-target hybrid Hybrid GAS109. Preferably, the hybridization chambers each have a wall portion that is transparent to the light emitted by the probe. GAS109. Preferably, the CMOS circuit is provided with the light sensor, wherein the wall portion is located between the probes and the light sensor. GAS109. Preferably, the wall portion is doped with a layer of cerium oxide. GAS109. Preferably, the probes each have an ECL luminophore that emits photons when excited by current between the electrodes and a functional portion for quenching photon emission from the ECL luminophore by resonance energy transfer. -103- 201211533 GAS 109. Preferably, the probes are configured such that when the probe forms a probe-target hybrid, the functional portion for quenching photon emission from the ECL luminophore is further derived from the ECL Luminous group. GAS 109. Preferably, the CMOS circuit is configured to provide electrical pulses to the electrodes. GAS 1 09. 13 period. Preferably, the electrical pulse has a GAS 1 09 of less than 0 · 69 seconds. 1 4 Preferably, the current of the electrical pulse is between 0. 1 nai to 69. 0 nai pei. GAS 109. Preferably, the electrodes have an anode and a cathode, each having a finger configuration, the finger structures being configured to intersect the fingers of the anode with the fingers of the cathode. GAS 1 09. 16 Preferably, the anode and the cathode are separated by 0. Dielectric gap of 4 microns to 2 microns wide. GAS 1 09. 1 7 microns. Preferably, the transparent wall portion has a thickness of less than 1,600 GAS 1 09. Preferably, the light sensor is positioned in an array of the photodiodes that are associated with the hybridization chamber such that each of the hybridization chambers corresponds to one of the photodiodes. GAS109. Preferably, each of the active regions is in the same plane, and the electrodes are patterned to form the conductive material of the separated anode and cathode, the plane of the layer and the photodiodes The planes of the active regions of the body are parallel. -104- 201211533 GAS109. Preferably, one of the electrode pairs causes oxidation or reduction of the luminophore to produce a working electrode of an excited species that emits photons, the working electrode being configured such that the probes are interposed The photodiode is between the working electrode and the working electrode. This high-angle emission collection provides a comprehensive solution that is both mass-produced and inexpensive, with low system components and provides a lightweight, highly mobile system. The large-angle emission of light increases the sensitivity of reading, which eliminates the need for optical components to be used in the string of light collecting elements. The detection and analysis of the target based on electrochemiluminescence eliminates the need for excitation light source, excitation optics and optics. The detection system of the filter element provides a lighter and less expensive detection system. It is not necessary to reject any excitation light and simplify the detector circuit, making the detection system even cheaper. GAS 1 1 0. 1 Aspects of the invention provide a microfluidic device for detecting a target molecule in a fluid, the microfluidic device comprising: an array of electrochemiluminescence (ECL) probes for reacting with the target φ molecules Forming a probe-target complex; an electrode for receiving electrical pulses, the probe-target composite system being configured to emit photons of light when excited by current between the electrodes: and for detecting A light sensor that emits light from the probe; wherein, when used, the fluid is added to the probes to prevent subsequent addition of other fluids to the probes. GAS 11 0. Preferably, the microfluidic device also has an array of chambers, 105-201211533, wherein each of the chambers includes a pair of electrodes and a probe for one of the targets, wherein the fluid is acted upon by capillary action Fill each of these rooms. GAS110. 3 Preferably, the volumes of the chambers are each less than 900,000 cubic microns. GAS 11 0. Preferably, the distance between the light sensor and the probe is less than 1,600 microns. GAS110. Preferably, the microfluidic device also has: a support substrate; a CMOS circuit for supplying electrical pulses to the electrodes; and a flow path containing the target fluid; wherein the CMOS is interposed between the chambers and the The substrates are supported, and the flow path attracts the fluid to each of the chambers by capillary action. GAS 110. Preferably, the flow system biological sample and the target nucleic acid sequence, each of the probes has a nucleic acid sequence complementary to the target and the chambers are used to hybridize the probes to form a probe-target hybrid Hybrid room. G. AS1 10. Preferably, the hybridization chambers each have a wall portion that is transparent to the light emitted by the probe. GAS110. Preferably, the CMOS circuit is provided with the light sensor, wherein the wall portion is located between the probes and the light sensor. GAS110. Preferably, the wall portion is doped with a layer of cerium oxide. GAS110. Preferably, the probes each have an ECL luminophore that emits photons when excited by current between the electrodes and a functional portion for quenching photon emission from the ECL luminophore by resonance energy transfer. 8 -106- 201211533 GAS 11 0. Preferably, the probes are configured such that when the probe forms a probe-target hybrid, the functional moiety for quenching photon emission from the ECL luminophore is further derived from the ECL Luminous group. GAS 110. Preferably, the C Μ O S circuit is configured to provide electrical pulses to the electrodes. GAS1 10. 13 Preferably, the electrical pulse has a value less than 0. φ period of 69 seconds. GAS 1 10. 14 Preferably, the current of the electrical pulse is between 0. 1 nai to 69. 0 nai pei. GAS110. Preferably, the electrodes have an anode and a cathode, each having a finger configuration, the finger structures being configured to intersect the fingers of the anode with the fingers of the cathode. GAS 1 10. 16 Preferably, the anode and the cathode are separated from each other. Dielectric gap of 4 microns to 2 microns wide. φ GAS1 10. Preferably, the transparent wall portion has a thickness of less than 1,600 microns. GAS1 10. Preferably, the light sensor is an array of photodiodes locating the hybridization chamber such that each of the hybridization chambers corresponds to one of the photodiodes. GAS110. Preferably, the photodiodes have planar active surface regions for receiving light from the luminophore, each of the active surface regions being in the same plane, and the layers of the electrodes are patterned. To form the conductive material of the separate anode and cathode, the plane of the layer -107-201211533 is parallel to the plane of the active surface region of the photodiode 〇GAS 11 0. Preferably, one of the electrode pairs causes oxidation or reduction of the luminophore to produce a working electrode of an excited species that emits photons, the working electrode being configured such that the probes are interposed The photodiode is between the working electrode and the working electrode. The easy-to-use, mass-produced, inexpensive and lightweight microfluidic device accepts a biological sample, uses its integrated imaging sensor to hybridize the probe to identify the nucleic acid sequence of the sample, and provides an electronic result on the output pad. The electrochemiluminescence-based detection analysis eliminates the need for detection systems that require excitation light sources, excitation optics, and optical filter elements to provide a lighter and less expensive inspection system. It is not necessary to reject any excitation light and simplify the detector circuit, making the detection system even cheaper. GAS 1 1 1 · 1 Aspects of the invention provide a wafer-on-lab (LOC) device for detecting a target molecule in a fluid, the LOC device comprising an array of electrochemiluminescence (ECL) probes for use in an array Reacting with the target molecules to form a probe-target complex: an electrode for receiving electrical pulses configured to emit photons of light when excited by current between the electrodes And a light sensor for detecting light emitted by the probe-target complex. GAS 1 11. 2 Preferably, the LOC device also has: -108-201211533 support substrate; a CMOS circuit for supplying electrical pulses to the electrodes; an array of chambers, each chamber containing a reaction for reacting with the target molecules to form a probe a probe of the target complex and a pair of electrodes; and a flow path of the fluid containing the target; wherein the CMOS is interposed between the chamber and the support substrate, and the flow path attracts the fluid by capillary action To each of these rooms. GAS111. Preferably, the flow system biological sample and the target nucleic acid sequence, each of the probes has a nucleic acid sequence complementary to the target and the chambers are used to hybridize the probes to form a probe-target hybrid Hybrid room. GASU1. Preferably, the hybridization chambers each have a wall portion that is transparent to the light emitted by the probe. GAS111. Preferably, the CMOS circuit is provided with the light sensor, wherein the wall portion is located between the probes and the light sensor. GAS 11 1. Preferably, the wall portion is doped with a layer of cerium oxide. GAS111. Preferably, the probes each have an ECL luminophore that emits photons when excited by current between the electrodes and a functional portion for quenching photon emission from the ECL luminophore by resonance energy transfer. 〇GAS 1 1 1. Preferably, the probes are configured such that when the probe forms a probe-target hybrid, the functional moiety for quenching photon emission from the ECL luminophore is further derived from the ECL Luminous group. GAS 1 11. Preferably, the CMOS circuit is configured to provide -109 - 201211533 electrical pulses to the electrodes. GAS111. 10 Preferably, the electrical pulse has a value less than 0. During the 69 seconds period. GAS111. 11 Preferably, the current of the electrical pulse is between 0. 1 nai to 69. 0 nai pei. GAS11 1. Preferably, the electrodes have an anode and a cathode, each having a finger configuration, the finger structures being configured to intersect the fingers of the anode with the fingers of the cathode. GAS111. 13 Preferably, the anode and the cathode are separated by 0. Dielectric gap of 4 microns to 2 microns wide. GAS111. Preferably, the transparent wall portion has a thickness of less than 1,600 microns. GAS1 11. Preferably, the light sensor is an array of photodiodes locating the hybridization chamber such that each of the hybridization chambers corresponds to one of the photodiodes. GAS1 11. Preferably, the photodiodes each have a planar active surface area such that the planar active surface regions collectively provide the collection surface, each of the active surface regions being in the same plane, and the electrode layers are Patterned to form a separate conductive material of the anode and cathode, the plane of the layer being parallel to the plane of the active surface area of the photodiodes. GAS 1 11. Preferably, one of the electrode pairs causes oxidation or reduction of the luminophore to produce a working electrode of an excited species that emits photons, the working electrode being configured such that the probes are interposed -110- 201211533 Between the photodiode and the working electrode. GAS1 11. Preferably, the luminophore is an organometallic complex. GAS 1 1 1 1 9 Preferably, the organometallic complex is an organogermanium complex molecule. GAS111. Preferably, the LOC device also has a polymerase interlocking reaction (PCR) portion for amplifying the target nucleic acid sequence in the sample. φ This easy-to-use, mass-produced, inexpensive and lightweight LOC device accepts a biological sample, uses its integrated imaging sensor to hybridize the probe to identify the nucleic acid sequence of the sample, and provides an electronic result on the output pad. The electrochemiluminescence-based detection analysis eliminates the need for detection systems that require excitation light sources, excitation optics, and optical filter elements to provide a lighter and less expensive inspection system. It is not necessary to reject any excitation light and simplify the detector circuit, making the detection system even cheaper. GAS1 12. 1 This aspect of the invention provides a wafer-on-lab (LOC) device for exciting an electrochemical luminescence group, the LOC device comprising: a support substrate; an array of electrochemiluminescence (ECL) probes, which is used Reacting with the target molecules to form a probe-target complex, each of which has a luminescent group for emitting photons in an excited state; an electrode for receiving an electrical pulse to utilize the electrodes The current between them excites the luminophores; and a CMOS circuit for controlling the electrical pulses transmitted to the electrodes; the CMOS circuit of -111 - 201211533 is interposed between the support substrate and the probe array. GAS112. 2 Preferably, the LOC device also has: a light sensor for detecting light emitted by the probe-target complex; an array of chambers, each chamber containing a reaction for reacting with the target molecules to form a probe a probe of the target complex and a pair of electrodes; and a flow path containing the fluid of the target; wherein the flow path attracts the fluid to each of the chambers by capillary action. GAS1 12. Preferably, the flow system biological sample and the target nucleic acid sequence, each of the probes has a nucleic acid sequence complementary to the target and the chambers are used to hybridize the probes to form a probe-target hybrid Hybrid room. GAS1 12. Preferably, the hybridization chambers each have a wall portion that is transparent to the light emitted by the probe. GAS112. Preferably, the CMOS circuit is provided with the light sensor, wherein the wall portion is located between the probes and the light sensor. GAS II 2. Preferably, the wall portion is doped with a layer of cerium oxide. GAS112. Preferably, each of the probes has a functional portion that is resonated by resonance energy to quench photon emission from the ECL luminophore. GAS112. Preferably, the probes are configured such that when the probe forms a probe-target hybrid, the functional moiety for quenching photon emission from the ECL luminophore is further derived from the ECL Luminous group. GAS1 12. Preferably, the CMOS circuit is configured to provide 8 - 112 201211533 electrical pulses to the electrodes. GAS1 12. 10 Preferably, the electrical pulse has a value less than 0. During the 69 seconds period. GAS112. Il preferably, the current of the electrical pulse is between 0. 1 nai to 69. 0 nai pei. GAS 11 2. Preferably, the electrodes have an anode and a cathode, each having a finger configuration, the finger structures being configured to intersect the fingers of the anode with the fingers of the cathode. GAS 1 12. 13 Preferably, the anode and the cathode are separated by 0. Dielectric gap of 4 microns to 2 microns wide" GAS 1 12. Preferably, the transparent wall portion has a thickness of less than 1,600 microns. GAS112. Preferably, the light sensor is an array of photodiodes locating the hybridization chamber such that each of the hybridization chambers corresponds to one of the photodiodes. GAS112. Preferably, the photodiodes each have a planar active surface area such that the planar active surface regions collectively provide the collection surface, each of the active surface regions being in the same plane, and the electrode layers are Patterned to form a separate conductive material of the anode and cathode, the plane of the layer being parallel to the plane of the active surface area of the photodiodes. GAS112. Preferably, one of the electrode pairs causes oxidation or reduction of the luminophore to produce a working electrode of an excited species that emits photons, the working electrode being configured such that the probes are interposed -113- 201211533 Between the photodiode and the working electrode. GAS1 12. Preferably, the luminophore is an organometallic complex. GAS 11 2. Preferably, the organometallic complex is an organogermanium complex molecule. GAS1 12. Preferably, the LOC device also has a polymerase interlocking reaction (PCR) portion for amplifying the target nucleic acid sequence in the sample. The integrated driver for exciting the electroluminescent luminophore on the LOC device operates from the universal USB, providing a system that is easy to use, mass-produced, inexpensive, and lightweight with a small number of components. The electrochemiluminescence-based detection analysis eliminates the need for detection systems that require excitation light sources, excitation optics, and optical filter elements to provide a lighter and less expensive inspection system. It is not necessary to reject any excitation light and simplify the detector circuit, making the detection system even cheaper. GAS1 13. 1 Aspects of the invention provide a wafer-on-lab (LOC) device for detecting a target molecule in a fluid, the LOC device comprising an array of electrochemiluminescence (ECL) probes for use with the target molecule Reacting to form a probe-target complex; and an electrode for receiving an electrical pulse; wherein the probes each have an ECL luminophore that emits photons when excited by current between the electrodes and is used for resonance energy Transfer quenching the functional portion of photon emission from the ECL luminophore. GAS113. Preferably, the LOC device also has: -114-201211533 A light sensor for detecting light emitted by the probe-target complex. GAS1 13. Preferably, the LOC device also has: a support substrate; a CMOS circuit for supplying electrical pulses to the electrodes; an array of chambers, each chamber containing a complex for reacting with the target molecules to form a probe-target complex a probe and a pair of electrodes; and φ a flow path containing the target fluid; wherein the CMOS is interposed between the chambers and the support substrate, and the flow path attracts the fluid to each by capillary action Waiting room. GAS113. Preferably, the flow system biological sample and the target nucleic acid sequence, each of the probes has a nucleic acid sequence complementary to the target and the chambers are used to hybridize the probes to form a probe-target hybrid Hybrid room. GAS113. Preferably, the hybridization chambers each have a wall portion that is transparent to the light emitted by the probe φ. GAS113. Preferably, the CMOS circuit is provided with the light sensor, wherein the wall portion is located between the probes and the light sensor. GAS 1 13. Preferably, the wall portion is doped with a layer of cerium oxide. GAS113. Preferably, the probes are configured such that when the probe forms a probe-target hybrid, the functional moiety for quenching photon emission from the ECL luminophore is further derived from the ECL Luminous group. GAS113. Preferably, the CMOS circuit is configured to provide electrical pulses to the electrodes. -115- 201211533 GASl 13. Preferably, the electrical pulse has a value less than 〇.  6 9 seconds period. GAS113. Il preferably, the current of the electrical pulse is between 0. 1 nai to 69. 0 nai pei. GAS1 13. Preferably, the electrodes have an anode and a cathode, each having a finger configuration, the finger structures being configured to intersect the fingers of the anode with the fingers of the cathode. GAS1 13. 13 Preferably, the anode and the cathode are separated by 0. Dielectric gap of 4 microns to 2 microns wide. GAS 113. Preferably, the transparent wall portion has a thickness of less than 1,600 microns. GAS 1 13. Preferably, the light sensor is an array of photodiodes locating the hybridization chamber such that each of the hybridization chambers corresponds to one of the photodiodes. GAS113. Preferably, the photodiodes each have a planar active surface area such that the planar active surface areas collectively provide the collecting surface, each of the active surface areas being in the same plane, and the electrodes are patterned by a layer A conductive material is formed to form the separate anode and cathode, the plane of the layer being parallel to the plane of the active surface area of the photodiodes. GAS113. Preferably, one of the electrode pairs causes oxidation or reduction of the luminophore to produce a working electrode of an excited species that emits photons, the working electrode being configured such that the probes are interposed The photodiode is between the working electrode and the working electrode. -116- 201211533 GASl 13. Preferably, the luminophore is an organometallic complex. GAS 11 3. Preferably, the organometallic complex is an organogermanium complex molecule. GAS113. Preferably, the LOC device also has a polymerase interlocking reaction (PCR) portion for amplifying the target nucleic acid sequence in the sample. The easy-to-use, mass-produced, inexpensive and lightweight LOC device accepts a biological sample, and uses its integrated imaging sensor to hybridize with an electrochemiluminescence resonance energy transfer probe to identify the nucleic acid sequence of the sample, and The output pad provides an electronic result, and the electrochemiluminescent resonance energy transfer probe provides a high specificity and high reliability for detecting the sequence of the target. The electrochemiluminescence-based detection analysis eliminates the need for detection systems that require excitation light sources, excitation optics, and optical filter elements to provide a lighter and less expensive inspection system. It is not necessary to reject any excitation light and simplify the detector circuit, making the detection system even cheaper. GAS 1 14. 1 This aspect of the invention provides a wafer-on-lab (LOC) device for detecting hybridization of a target nucleic acid sequence, the LOC device comprising: electrochemiluminescence (ECL), resonance energy transfer, attachment to a primer, and stem-loop detection Needle, its system is used with. The labeled nucleic acid sequences hybridize to form probe-target hybrids, each of which has a loop portion containing the target nucleic acid sequence, and an primer for extending along the target nucleic acid sequence to form a nucleic acid sequence complementary to the target An ECL luminophore for emitting photons in an excited state and a functional portion for quenching photons from the ECL luminophore by resonant energy transfer - 117 - 201211533; and for receiving electrical pulses to excite such An electrode of an ECL luminophore; wherein forming the complementary nucleic acid sequence when used causes the loop portion to open such that the target nucleic acid sequence hybridizes to the complementary nucleic acid sequence and the ECL luminophore is moved away from the functional portion. GAS 1 14. Preferably, the LOC device also has: a light sensor for detecting light emitted when the ECL luminophore is in the probe-target hybrid configuration. GAS 1 14. Preferably, the LOC device also has: a support substrate; a CMOS circuit for supplying electrical pulses to the electrodes; a hybrid chamber containing a probe for hybridizing with the target and a pair of electrodes: and containing the same a flow path of the target fluid; wherein the CMOS is interposed between the hybridization chamber and the support substrate, and the flow path attracts the fluid to each of the hybridization chambers by capillary action. GAS114. Preferably, the LOC device also has a lysis unit, wherein the flow system contains a biological sample of the cell and the lysate destroys the cell membrane to release any genetic material therein. GAS114. Preferably, the hybridization chambers each have a wall portion that is transparent to the light emitted by the probe. GAS1 14. Preferably, the CMOS circuit is provided with the light sensor, wherein the wall portion is located between the probes and the light sensor. 8 -118- 201211533 GAS 11 4. Preferably, the wall portion is doped with a layer of cerium oxide. GAS114. Preferably, the probes are configured such that when the probe forms a probe-target hybrid, the functional moiety for quenching photon emission from the ECL luminophore is further derived from the ECL Luminous group. GAS 114. Preferably, the CMOS circuit is configured to provide electrical pulses to the electrodes. GAS114. 10 Preferably, the electrical pulse has a value less than 0. During the 69 seconds period. GAS 11 4. 11 Preferably, the current of the electrical pulse is between 0. 1 nai to 69. 0 nai pei. GAS 11 4. Preferably, the electrodes have an anode and a cathode, each having a finger configuration, the finger structures being configured to intersect the fingers of the anode with the fingers of the cathode. GAS 1 14. 13 Preferably, the anode and the cathode are separated from each other. Dielectric gap of 4 microns to 2 microns wide. GAS1 14. Preferably, the transparent wall portion has a thickness of less than 1,600 microns. GAS114. Preferably, the light sensor is an array of photodiodes locating the hybridization chamber such that each of the hybridization chambers corresponds to one of the photodiodes. GAS114. Preferably, the photodiodes each have a planar active surface area such that the planar active surface regions collectively provide the collection surface, each of the active surface regions being in the same plane 'and the layers are patterned Patterned to form the separate anode and cathode leads -119 - 201211533 electrical material, the plane of the layer being parallel to the plane of the active surface area of the photodiodes. GAS114. Preferably, one of the electrode pairs causes oxidation or reduction of the luminophore to produce a working electrode of an excited species that emits photons, the working electrode being configured such that the probes are interposed The photodiode is between the working electrode and the working electrode. GAS1 14. Preferably, the luminophore is an organometallic complex. GAS114. Preferably, the organometallic complex is an organogermanium complex molecule. GAS1 14. Preferably, the LOC device also has a polymerase interlocking reaction (PCR) portion for amplifying the target nucleic acid sequence in the sample. The easy-to-use, mass-produced, inexpensive and lightweight LOC device accepts a biological sample, amplifies the nucleic acid target in the sample, and uses the integrated imaging sensor to connect the stem ring to the electrochemiluminescence resonance energy transfer primer. The probe hybridizes to identify the nucleic acid sequence of the sample, and provides an electronic result at the output pad, and the stem-loop probe attached to the primer provides a plurality of optimal parallel amplification reactions to be performed, and also provides high specificity, Sensitive and reliable detection of the target sequence. The electrochemiluminescence-based detection analysis eliminates the need for detection systems that require excitation light sources, excitation optics, and optical filter elements to provide a lighter and less expensive inspection system. It is not necessary to reject any excitation light and simplify the detector circuit, making the detection system even cheaper. GAS 11 5 . 1 This aspect of the invention provides a wafer-on-lab (LOC) device for detecting hybridization of a target -120 - 201211533 nucleic acid sequence, the LOC device comprising: electrochemiluminescence (ECL), resonance energy transfer, and primer connection a linear probe for hybridizing to the target nucleic acid sequence to form a probe-target hybrid, each of the probes having a linear portion comprising the target nucleic acid sequence for extension along the target nucleic acid sequence An primer for forming a nucleic acid sequence complementary to the target, an ECL luminophore for emitting photons in an excited state, and a functional portion for quenching photon emission from the ECL luminophore by resonance energy transfer; and for accepting Electrically pulsing to excite the electrodes of the ECL luminophores; wherein, when used, replicating the target nucleic acid sequence causes the linear portion to dissociate from the functional portion such that the complementary nucleic acid sequence within it hybridizes to the target nucleic acid sequence, and The photons emitted by the ECL luminophore are not quenched. GAS115. 2 Preferably, the LOC device also has: a light sensor for detecting light emitted by the ECL luminophore. GAS 1 15. Preferably, the LOC device also has: a support substrate; a CMOS circuit for supplying electrical pulses to the electrodes; a hybrid chamber containing a probe for hybridizing with the target and a pair of electrodes: and containing the same a flow path of the target fluid; wherein the CMOS is interposed between the hybridization chamber and the support substrate, and the flow path attracts the fluid to each of the hybridization chambers by capillary action. -121 - 201211533 GASl 15. Preferably, the LOC device also has a lysis unit, wherein the flow system contains a biological sample of the cell and the lysate destroys the cell membrane to release any genetic material therein. GAS 1 15. Preferably, the hybridization chambers each have a wall portion that is transparent to the light emitted by the probe. GAS115. Preferably, the CMOS circuit is provided with the light sensor, wherein the wall portion is located between the probes and the light sensor. GAS115. Preferably, the wall portion is doped with a layer of cerium oxide. GAS 1 1 5. Preferably, the probes are configured such that when the probe forms a probe-target hybrid, the functional moiety for quenching photon emission from the ECL luminophore is further derived from the ECL Luminous group. GAS 1 15. Preferably, the C Μ O S circuit is configured to provide electrical pulses to the electrodes. GAS 11 5. 10 Preferably, the electrical pulse has a value less than 0. During the 69 seconds period. GAS115. Il preferably, the current of the electrical pulse is between 〇. 1 nai to 6 0 nai pei. GAS 11 5. Preferably, the electrodes have an anode and a cathode, each having a finger configuration, the finger structures being configured to intersect the fingers of the anode with the fingers of the cathode. GAS 115. 13 Preferably, the anode and the cathode are separated from each other.  Dielectric gap of 4 microns to 2 microns wide. GAS 1 15. Preferably, the transparent wall portion has a thickness of less than 1,6 Å. -122- 201211533 GAS 115. Preferably, the light sensor is an array of photodiodes locating the hybridization chamber such that each of the hybridization chambers corresponds to one of the photodiodes. GAS 115. Preferably, the photodiodes each have a planar active surface area such that the planar active surface areas collectively provide the collecting surface, each of the active surface areas being in the same plane, and the electrodes are patterned by a layer Patterned to form the separate anode and cathode conductive material, the plane of the layer being parallel to the plane of the active surface area of the photodiodes. GAS1 15. Preferably, one of the electrode pairs causes oxidation or reduction of the luminophore to produce a working electrode of an excited species that emits photons, the working electrode being configured such that the probes are interposed The photodiode is between the working electrode and the working electrode. GAS 11 5.  Preferably, the luminophore is an organometallic complex. GAS 1 1 5 .  Preferably, the organometallic complex is an organogermanium compound molecule. GAS1 15. Preferably, the LOC device also has a polymerase interlocking reaction (PCR) portion for amplifying the target nucleic acid sequence ' in the sample. The easy-to-use, mass-produced, inexpensive and lightweight LOC device accepts a biological sample, amplifies the nucleic acid target in the sample, and utilizes an integrated imaging sensor to linearly link with an electrochemiluminescence resonance energy transfer primer. Needle hybridization to identify the nucleic acid sequence of the sample, and to provide an electronic result at the output pad, the linear probe coupled to the primer provides a large number of optimal parallel amplification reactions to be performed by -123-201211533, also providing high specificity Detection of the sequence of the target for sex, sensitivity and reliability. The electrochemiluminescence-based detection analysis eliminates the need for detection systems that require excitation light sources, excitation optics, and optical filter elements to provide a lighter and less expensive inspection system. It is not necessary to reject any excitation light and simplify the detector circuit, making the detection system even cheaper. GAS 1 17. 1 This aspect of the invention provides a wafer-on-lab (LOC) device for amplifying and detecting a target nucleic acid sequence, the LOC device comprising: electrochemiluminescence (ECL) for hybridization with the target nucleic acid sequence, Resonance energy transfer, stem-loop probes, each of which has a loop portion containing a sequence complementary to the target nucleic acid sequence, an ECL luminophore for emitting photons in an excited state, for quenching by resonance energy transfer a functional moiety derived from photon emission of the ECL luminophore, and a covalently linked primer for extending along a complementary sequence denatured from the target nucleic acid sequence to replicate the target nucleic acid sequence; for heating to circulate the target nucleic acid sequence for performing a polymerase chain reaction (PCR) heater, wherein the covalently linked primer is bonded to an oligonucleotide comprising the target nucleic acid sequence; and an electrode for receiving an electrical pulse to excite the ECL luminophore; wherein when used Copying the target nucleic acid sequence causes the loop portion to be opened such that the complementary nucleic acid sequence therein hybridizes to the target nucleic acid sequence and the ECL luminophore is moved to From the functional moiety. -124- 201211533 GASl 17. 2 Preferably, the LOC device also has: a light sensor for detecting light emitted by the ECL luminophore. GAS 1 17. Preferably, the LOC device also has: a support substrate; a CMOS circuit for activating the heaters and supplying electrical pulses to the electrodes; and a probe for hybridizing with the targets, the heaters a hybridization chamber of one and a pair of electrodes; and a flow path of the fluid containing the target; wherein the CMOS circuit is interposed between the hybridization chamber and the support substrate, and the flow path is capillary The action draws the fluid to each of the hybrid chambers. GAS117. Preferably, the LOC device also has a lysis unit, wherein the flow system contains a biological sample of the cell and the lysate destroys the cell membrane to release any genetic material therein. GAS 117. Preferably, the hybridization chambers each have a wall portion that is transparent to the light emitted by the probe φ. GAS 11 7. Preferably, the CMOS circuit is provided with the light sensor, wherein the wall portion is located between the probes and the light sensor. GAS 1 1 7. Preferably, the wall portion is doped with a layer of cerium oxide. GAS 1 1 7. Preferably, the covalently linked primer is attached to the functional moiety for quenching photon emission from the ECL luminophore and the loop moiety is interposed between the ECL luminophore and the functional moiety. GAS 11 7. Preferably, the CMOS circuit is configured to provide electrical pulses to the electrodes. -125- 201211533 GASl 17. Preferably, the electrical pulse has a period of less than 〇·69 seconds. GAS117. Il preferably, the current of the electrical pulse is between 〇. 1 nai to 69. 0 nai pei. GAS 11 7.  Preferably, the electrodes have an anode and a cathode, each having a finger configuration, the finger structures being configured to intersect the fingers of the anode with the fingers of the cathode. GAS117. 13 Preferably, the anode and the cathode are separated by 0. Dielectric gap of 4 microns to 2 microns wide. GAS117. Preferably, the transparent wall portion has a thickness of less than 1,600 microns. GAS 11 7. Preferably, the light sensor is an array of photodiodes locating the hybridization chamber such that each of the hybridization chambers corresponds to one of the photodiodes. GAS 11 7.  Preferably, the photodiodes each have a planar active surface area such that the planar active surface areas collectively provide the collecting surface, each of the active surface areas being in the same plane, and the electrodes are patterned by a layer Patterned to form the conductive material of the separate anode and cathode, the plane of the layer being parallel to the plane of the active surface area of the photodiodes. GAS117. Preferably, one of the electrode pairs causes oxidation or reduction of the luminophore to produce a working electrode of an excited species that emits photons. The position of the working electrode is configured such that the probes are interposed The photodiode is between the working electrode and the working electrode. -126- 201211533 GAS117. Preferably, the luminophore is an organometallic complex. GAS 11 7. Preferably, the organometallic complex is an organogermanium complex molecule. GAS117. Preferably, the LOC device also has a dialysis section for diverting cells smaller than a predetermined size valve to separate streams. The easy-to-use, mass-produced, inexpensive and lightweight LOC device accepts a biological sample, amplifies the nucleic acid target in the sample, and uses the integrated φ-type imaging sensor to connect to the stem via an electrochemiluminescence resonance energy transfer primer. Loop probe hybridization to identify the nucleic acid sequence of the sample and provide an electronic result at the output pad, the stem-loop probe attached to the primer provides a large number of optimal parallel amplification reactions to be performed, and also provides high specificity Detection of Sensitivity and Reliability The Sequence of Measurements This electrochemiluminescence-based detection analysis eliminates the need for detection systems that require excitation, excitation, and optical filter components to provide a lighter and less expensive inspection system. It is not necessary to reject any excitation light and simply φ the detector circuit, making the detection system even cheaper. GAS 1 18. 1 This aspect of the invention provides a wafer-on-lab (LOC) device for amplifying and detecting a target nucleic acid sequence, the LOC device comprising: electrochemiluminescence (ECL) for hybridization with the target nucleic acid sequence, Resonance energy transfer, linear probes, each of which has a linear portion comprising a sequence complementary to the target nucleic acid sequence, an ECL luminophore for emitting photons in an excited state, for quenching by resonance energy transfer from a functional portion of the photon emission of the ECL luminophore, and a covalently linked primer for extending along the complementary sequence denaturing from the nucleic acid sequence of the standard -127-201211533 to replicate the target nucleic acid sequence: for heating the loop a nucleic acid sequence for performing a polymerase chain reaction (PCR) heater, wherein the covalently linked primer is bonded to an oligonucleotide comprising the target nucleic acid sequence: and an electrode for receiving an electrical pulse to excite the ECL luminophore; Wherein, when used, the replication of the target nucleic acid sequence causes the linear portion to dissociate from the functional portion such that the complementary nucleic acid sequence within it and the target nucleus Sequences hybridize, and the photons emitted by the ECL luminophore which is not quenched. GAS118. 2 Preferably, the LOC device also has: a light sensor for detecting light emitted by the ECL luminophore. GAS1 18. Preferably, the LOC device also has: a support substrate; a CMOS circuit for activating the heaters and supplying electrical pulses to the electrodes; and a probe for hybridizing with the targets, the heaters a hybridization chamber of at least one of the electrodes and a pair of electrodes; and a flow path of the fluid containing the target; wherein the CMOS is interposed between the hybridization chamber and the support substrate, and the flow path attracts the capillary by capillary action Fluid to each of the hybridization chambers. GAS1 18. Preferably, the LOC device also has a lysis unit, wherein the flow system contains a biological sample of the cell and the lysate destroys the cell membrane to release any genetic material therein. -128 - 201211533 GAS118. Preferably, the hybridization chambers each have a wall portion that is transparent to the light emitted by the probe. GAS118. Preferably, the CMOS circuit is provided with the light sensor, wherein the wall portion is located between the probes and the light sensor. GAS118. Preferably, the wall is provided with a layer of cerium oxide. GAS 118. Preferably, the probe has a PCR blocker group between the covalently linked primer and the linear portion. Φ GAS118. Preferably, the CMOS circuit is configured to provide electrical pulses to the electrodes. GAS1 18. 10 Preferably, the electrical pulse has a value less than 〇.  6 9 seconds period. GAS118. Il preferably, the current of the electrical pulse is between 〇.  1 nai to 69. 0 nai pei. GAS 1 1 8 . Preferably, the electrodes have an anode and a cathode each having a finger-like configuration, the finger structures being configured to intersect the finger of the anode φ with the finger of the cathode. GAS 11 8. 13 Preferably, the anode and the cathode are separated by 0. Dielectric gap of 4 microns to 2 microns wide. GAS 118. Preferably, the transparent wall portion has a thickness of less than 1,600 microns. GAS 1 18. Preferably, the light sensor is an array of photodiodes locating the hybridization chamber such that each of the hybridization chambers corresponds to one of the photodiodes. GAS118. Preferably, the photodiodes each have a planar main-129-201211533 moving surface area such that the planar active surface areas collectively provide the collecting surface, each of the active surface areas being in the same plane, and The electrode system layer is patterned to form the conductive material of the separate anode and cathode, the plane of the layer being parallel to the plane of the active surface area of the photodiodes. GAS 1 18. Preferably, one of the electrode pairs causes oxidation or reduction of the luminophore to produce a working electrode of an excited species that emits photons, the working electrode being configured such that the probes are interposed The photodiode is between the working electrode and the working electrode. GAS118. Preferably, the luminophore is an organometallic complex. GAS118. Preferably, the organometallic complex is an organic ruthenium complex molecule GAS1 18. Preferably, the LOC device also has a dialysis section for diverting cells smaller than a predetermined size valve to separate streams. The easy-to-use, mass-produced, inexpensive and lightweight LOC device accepts a biological sample, amplifies the nucleic acid target in the sample, and utilizes an integrated imaging sensor to linearly link with an electrochemiluminescence resonance energy transfer primer. Needle hybridization to identify the nucleic acid sequence of the sample, and to provide an electronic result at the output pad, the linear probe coupled to the primer provides a large number of optimal parallel amplification reactions to be performed, also providing high specificity, sensitivity And the reliability of the detection of the sequence of the target. The electrochemiluminescence-based detection analysis eliminates the need for detection systems that require excitation light sources, excitation optics, and optical filter elements to provide a lighter and less expensive inspection system. There is no need to reject any excitation light. -130-201211533 This detector circuit makes the detection system even cheaper. GAS119. 1 This aspect of the invention provides a wafer-on-lab (LOC) device for amplifying and detecting a target nucleic acid sequence, the LOC device comprising: for limiting (digesting) double-stranded oligonucleosides at known junction sites a restriction enzyme for acid; a linker molecule for ligation to the junction of the double-stranded oligonucleotide; a heater for heating and circulating the oligonucleotide during a polymerase chain reaction (PCR); a primer for binding to a linker molecule on a single strand of the oligonucleotide after denaturation; a deoxyribonucleoside triphosphate (dNTP) for extending the primer along the single-stranded oligonucleotide; and for receiving electricity A pulse is applied to excite the electrodes of the ECL luminophores. GAS1 19. 2 Preferably, the LOC device also has: a light sensor for detecting light emitted by the ECL luminophore. GAS119. Preferably, the LOC device also has: a support substrate; a CMOS circuit for activating the heaters and supplying electrical pulses to the electrodes; and a probe for hybridizing with the targets, the heaters a hybridization chamber of at least one of the electrodes and a pair of electrodes; and a flow path of the fluid containing the target; wherein the CMOS circuit is interposed between the hybridization chamber and the support substrate, -131 - 201211533 and the flow path is borrowed The fluid is attracted to each of the hybridization chambers by capillary action. GAS 119. Preferably, the LOC device also has a lysis unit, wherein the flow system contains a biological sample of the cell and the lysate destroys the cell membrane to release any genetic material therein. GAS 1 1 9. 5 Preferably, the hybridization chambers each have a wall portion that can be penetrated by the light emitted by the probe. GAS119. 6 Preferably, the CMOS circuit has the light sensor, wherein the wall portion is located between the probe and the light sensor β GAS 11 9. Preferably, the wall portion is doped with a layer of cerium oxide. GAS119. Preferably, the probe has a stem-loop structure, the loop portion of which contains a sequence complementary to the target nucleic acid sequence, the loop portion being located in the functional portion for quenching photon emission from the ECL luminophore Between the ECL luminophores, hybridization with the target nucleic acid sequence opens the loop portion and moves the ECL luminophore away from the functional portion. GAS 1 19. Preferably, the CMOS circuit is configured to provide electrical pulses to the electrodes. GAS 1 19. 10 Preferably, the electrical pulse has a value less than 0. 69 seconds period" GAS119. Il preferably, the current of the electrical pulse is between 0. 1 nai to 69. 0 nai pei. GAS1 19. Preferably, the electrodes have an anode and a cathode, each having a finger configuration, the finger structures being configured to intersect the fingers of the anode with the fingers of the cathode. GAS 1 19. Preferably, the anode and the cathode are separated by a dielectric gap of -4 -132 - 201211533 μm to 2 μm wide. GAS119. Preferably, the transparent wall portion has a thickness of less than 1,600 microns. GAS119. Preferably, the light sensor is an array of photodiodes locating the hybridization chamber such that each of the hybridization chambers corresponds to one of the photodiodes. GAS119. Preferably, the photodiodes each have a planar main φ moving surface region such that the planar active surface regions collectively provide the collecting surface, each of the active surface regions being in the same plane, and the electrode systems are The layer is patterned to form the electrically conductive material of the separate anode and cathode, the plane of the layer being parallel to the plane of the active surface area of the photodiodes. GAS119. Preferably, one of the electrode pairs causes oxidation or reduction of the luminophore to produce a working electrode of an excited species that emits photons, the working electrode being configured such that the probes are interposed φ between the photodiode and the working electrode. GAS 119. Preferably, the luminophore is an organometallic complex. GAS 11 9.  Preferably, the organometallic complex is an organogermanium complex molecule. GAS119. Preferably, the LOC device also has a dialysis section for diverting cells smaller than a predetermined size valve to separate streams. The easy-to-use, mass-produced, inexpensive and lightweight L〇C device accepts a biological sample, amplifies the nucleic acid target in the sample, and uses the integrated imaging sensor to identify the nucleic acid sequence of the sample via probe hybridization. And at -133-201211533, the output pad provides electronic results, which provides the ability to genomic-level amplification and splitting by the adaptor primer. The electrochemiluminescence-based detection analysis eliminates the need for detection systems that require excitation light sources, excitation optics, and optical filter elements to provide a lighter and less expensive inspection system. It is not necessary to reject any excitation light and simplify the detector circuit, making the detection system even cheaper. GAS 120. 1 This aspect of the invention provides a wafer-on-lab (LOC) device for detecting a target nucleic acid sequence in a fluid, the LOC device comprising: electrochemiluminescence (ECL), resonance for hybridization with the target nucleic acid sequence Energy transfer probes, each of which contains a sequence complementary to the target nucleic acid sequence and an ECL luminophore for emitting photons in an excited state, for quenching photon emission from the ECL luminophore by resonance energy transfer Functional portion; an electrode for receiving an electrical pulse to excite the ECL luminophore; a hybrid chamber containing a probe for hybridization with the label, at least one of the heaters, and a pair of electrodes; a reagent reservoir containing reagents for addition to the fluid; wherein the hybrid chambers each have a volume of less than 900,000 cubic microns and the reagent reservoir has a volume of less than 1, 〇〇〇, 〇〇〇, 〇〇〇 cubic microns volume. GAS120. Preferably, the hybrid chambers each have a volume of less than 200,000 cubic microns and the reagent reservoir has a volume of less than 300,000,000 cubic microns. GAS 1 20. Preferably, the hybrid chambers each have a volume of less than 40,000 - 134 - 201211533 cubic microns and the reagent reservoir has a volume of less than 70,000,000 cubic microns. GAS120. Preferably, the hybrid chambers each have a volume of less than 9,000 cubic microns and the reagent reservoir has a volume of less than 20,000,000 cubic micrometers. GAS120. Preferably, the LOC device also has: a light sensor for detecting light emitted by the ECL luminophore. φ GAS 1 20. Preferably, the LOC device also has: a support substrate; a CMOS circuit for supplying electrical pulses to the electrodes; and a flow path containing the target fluid; wherein the CMOS circuit is interposed between the hybrid chambers Between the support substrates, the flow path attracts the fluid to each of the hybridization chambers by capillary action. GAS120. Preferably, the LOC device also has a lysis unit, wherein the flow system contains a biological sample of the cell and the lysate destroys the cell φ membrane to release any genetic material therein. GAS 12 0. Preferably, the hybridization chambers each have a wall portion that is transparent to the light emitted by the probe. GAS120. Preferably, the CMOS circuit is provided with the light sensor, wherein the wall portion is located between the probes and the light sensor. GAS 12 0. Preferably, the wall portion is doped with a layer of cerium oxide. GAS120. Il preferably, the probe has a stem-loop structure, the loop portion of which contains a sequence complementary to the target nucleic acid sequence, the loop portion being located in the functional portion for quenching photon emission from the ECL luminophore -135- 201211533 ECL luminophores, such that hybridization with the target nucleic acid sequence opens the loop portion and leaves the ECL luminophore away from the functional portion. GAS120. Preferably, the CMOS circuit is configured to provide electrical pulses to the electrodes. GAS 120. 13 Preferably, the electrical pulse has a value less than 0. During the 69 seconds period. GAS 120. 14 Preferably, the current of the electrical pulse is between 0. 1 nai to 69. 0 Nai'an Pei GAS 120. Preferably, the electrodes have an anode and a cathode each having a finger-like configuration, the finger structures being configured to intersect the fingers of the anode with the fingers of the cathode. GAS 120. 16 Preferably, the anode and the cathode are separated by 0. Dielectric gap of 4 microns to 2 microns wide. GAS120. Preferably, the transparent wall portion has a thickness of less than 1,600 microns. GAS 120. Preferably, the light sensor is an array of photodiodes locating the hybridization chamber such that each of the hybridization chambers corresponds to one of the photodiodes. GAS120. Preferably, the photodiodes each have a planar active surface area such that the planar active surface regions collectively provide the collection surface, each of the active surface regions being in the same plane, and the electrode layers are Patterned to form a separate conductive material of the anode and cathode, the plane of the layer being parallel to the plane of the active surface area of the photodiodes. -136- 201211533 GAS 1 20. Preferably, one of the electrode pairs causes oxidation or reduction of the luminophore to produce a working electrode of an excited species that emits photons, the working electrode being configured such that the probes are interposed The photodiode is between the working electrode and the working electrode. The low volume hybridization chamber and reagent reservoir represent, to some extent, a low probe and reagent volume, thereby providing a low probe and reagent cost and an inexpensive detection system. φ This electrochemiluminescence-based detection analysis eliminates the need for detection systems that require excitation light, excitation optics, and optical filter components to provide a lighter and less expensive inspection system. It is not necessary to reject any excitation light and simplify the detector circuit, making the detection system even cheaper. GAS 1 2 1 · 1 This aspect of the invention provides a wafer-on-lab (LOC) device for detecting a target nucleic acid sequence in a fluid, the LOC device comprising: electrochemiluminescence for detecting the target nucleic acid sequence ( ECL) probes, φ each of which has an ECL luminophore for emitting photons in an excited state, a functional moiety for quenching photon emission from the ECL luminophore by resonance energy transfer; at least one positive a control probe having one of the ECL luminophores but without the functional moiety for quenching photon emission; at least one negative control probe 'which does not have one of the ECL luminophores; And electrodes for receiving electrical pulses to excite the ECL luminophores. GAS121. 2 Preferably, the LOC device also has a light sensor located adjacent to the -137-201211533 probe for sensing those probes that respond to electrical pulses to produce photons. GAS121. Preferably, the LOC device also has a support substrate, wherein the light sensor is a charge coupled device (CCD) array between the probe and the support substrate. GAS 121. Preferably, the LOC device also has a support substrate, wherein the light sensor is an array of photodiodes on the support substrate that are registered with the probes. GAS121. Preferably, the photodiode array is less than 1,600 microns from the probe. GAS 121. Preferably, the LOC device also has a CMOS circuit on the support substrate, the array of photodiodes being a component of the CMOS circuit, wherein when used, the CMOS circuit response cannot sense from the positive control The EC L signal of the needle or response senses a signal from the negative control probe to cause an error signal. GAS 121 . Preferably, the LOC device also has a hybridization chamber comprising the probes and a pair of electrodes. GAS121. Preferably, the LOC device also has: a flow path containing the target fluid; wherein the CMOS circuit is interposed between the hybridization chamber and the support substrate, and the flow path attracts the fluid by capillary action To each of these hybrid chambers. GAS121. Preferably, the LOC device also has a lysis unit, wherein the flow system contains a biological sample of the cell and the lysate destroys the cell membrane to release any genetic material therein. -138- 201211533 GAS121. Preferably, the hybridization chambers each have a wall portion that is transparent to the light emitted by the probe. GAS121. Il Preferably, the CMOS circuit is provided with the light sensor, wherein the wall portion is located between the probes and the light sensor. GAS 12 1. 1 2 Preferably, the wall portion is doped with a layer of cerium oxide. GAS 12 1. Preferably, the probe has a stem-loop structure, the loop portion of which contains a sequence complementary to the target nucleic acid sequence, the loop portion being located at the functional portion of the φ for quenching photon emission from the ECL luminophore Between the ECL luminophore and the nucleic acid sequence of the target, the loop portion is opened and the ECL luminophore is moved away from the functional portion. GAS121. Preferably, the CMOS circuit is configured to provide electrical pulses to the electrodes. GAS 12 1. 1 5 Preferably, the electrical pulse has a value less than 0. During the 69 seconds period. GAS 121 .  Preferably, the current of the electrical pulse is between 0. 1 奈 amp to 69. 0 nai pei. GAS 1 2 1 .  Preferably, the electrodes have an anode and a cathode, each having a finger configuration, the finger structures being configured to intersect the fingers of the anode with the fingers of the cathode. GAS 12 1. 1 8 Preferably, the anode and the cathode are separated by 0. Dielectric gap of 4 microns to 2 microns wide. GAS121. Preferably, the transparent wall portion has a thickness of less than 1,600 microns. - GAS 12 1. Preferably, the LOC device also has an upper cover, and the -139-201211533 upper cover is provided with a reagent reservoir for adding to the sample before detecting the target sequence, wherein the electrodes and the probes are interposed Between the upper cover and the C Μ 0 S circuit. The hybridization array provides analysis of the target via hybridization and utilizes the control probe to improve the reliability of the analytical results. The electrochemiluminescence-based detection analysis eliminates the need for detection systems that require excitation light sources, excitation optics, and optical filter elements to provide a lighter and less expensive inspection system. It is not necessary to reject any excitation light and simplify the detector circuit, making the detection system even cheaper. GAS 1 22. 1 This aspect of the invention provides a wafer-on-lab (LOC) device for detecting a target nucleic acid sequence in a fluid, the LOC device comprising: an electrochemiluminescent (ECL) probe for detecting the target nucleic acid sequence, Each of the probes has an ECL luminophore for emitting photons in an excited state, a functional moiety for quenching photon emission from the ECL luminophore by resonance energy transfer; at least one positive control probe, For detecting nucleic acid sequences known to be present in the fluid; and electrodes for receiving electrical pulses to excite the ECL luminophores. GAS122. Preferably, the LOC device also has a light sensor located adjacent to the probe for sensing those probes that respond to electrical pulses to produce ECL photons. GAS122. Preferably, the LOC device also has a support substrate, wherein the light sensor is a charge-140-201211533 coupling device (CCD) array between the probe and the support substrate. GAS 122·4 Preferably, the LOC device also has a support substrate ‘where the light sensor is located on the support substrate and an array of photodiodes registered with the probes. GAS122. Preferably, the photodiode array is less than 1,600 microns from the probe. GAS122. Preferably, the LOC device also has a CMOS circuit on the φ substrate, the array of photodiodes being a component of the CMOS circuit, wherein when used, the CMOS circuit responds to the undetected from the positive control. The ECL of the probe emits an error signal. GAS122. Preferably, the LOC device also has a hybridization chamber comprising the probes and a pair of electrodes. GAS122. Preferably, the LOC device also has: a flow path containing the target fluid; wherein the CMOS circuit is interposed between the hybridization chamber and the support substrate, φ and the flow path attracts the capillary by capillary action Fluid to each of the hybridization chambers. GAS122. Preferably, the LOC device also has a lysis unit, wherein the flow system contains a biological sample of the cell and the lysate destroys the cell membrane to release any genetic material therein. GAS122. Preferably, the hybridization chambers each have a wall portion that is transparent to the light emitted by the probe. GAS122. Il Preferably, the CMOS circuit is provided with the light sensor, wherein the wall portion is located between the probes and the light sensor. GAS 122. Preferably, the wall portion is doped with a layer of cerium oxide. -141 - 201211533 GAS 122. Preferably, the probe has a stem-loop structure, the loop portion of which contains a sequence complementary to the target nucleic acid sequence, the loop portion being located in the functional portion for quenching photon emission from the ECL luminophore Between the ECL luminophores, hybridization with the target nucleic acid sequence opens the loop portion and moves the ECL luminophore away from the functional portion. GAS 1 22. Preferably, the C Μ Ο S circuit is configured to provide electrical pulses to the electrodes. GAS 1 22.  1 5 Preferably, the electrical pulse has a value less than 0. During the 69 seconds period. GAS 1 22. 1 6 Preferably, the current of the electrical pulse is between 0. 1 nai to 69. 0 nai pei. GAS 1 22. Preferably, the electrodes have an anode and a cathode, each having a finger configuration, the finger structures being configured to intersect the fingers of the anode with the fingers of the cathode. GAS 1 22. 1 8 Preferably, the anode and the cathode are separated by 0. Dielectric gap of 4 microns to 2 microns wide. GAS122. Preferably, the LOC device also has a PCR portion for amplifying the target nucleic acid sequence. GAS 1 22. Preferably, the LOC device also has an upper cover, the upper cover being provided with a reagent reservoir for adding to the sample before detecting the target sequence, wherein the electrodes and the probes are interposed between the upper cover and the Between CMOS circuits. The hybridization array provides analysis of the target via hybridization and utilizes the control probe to improve the reliability of the analytical results. -142- 201211533 This electrochemiluminescence-based detection analysis eliminates the need for detection systems that require excitation light sources, excitation optics, and optical filter elements to provide a lighter and less expensive inspection system. It is not necessary to reject any excitation light and simplify the detector circuit, making the detection system even cheaper. GAS 1 23 · 1 This aspect of the invention provides a wafer-on-lab (LOC) device for detecting a target nucleic acid sequence in a fluid, the LOC device comprising: φ for detecting electrochemiluminescence of the target nucleic acid sequence ( ECL) probes, each of which has an ECL luminophore for emitting photons in an excited state, a functional moiety for quenching photon emission from the ECL luminophore by resonance energy transfer; at least one positive control a probe having the ECL luminophore but having no functional portion for quenching photon emission; and an electrode for receiving electrical pulses to excite the ECL luminophores. GAS123. Preferably, the LOC device also has a light sensor located adjacent to the probe φ for sensing those probes that respond to electrical pulses to produce ECL photons. GAS123. 3 Preferably, the LOC device also has a support substrate in which the light is sensed. The device is a charge coupled device (CCD) array between the probe and the support substrate. GAS123. Preferably, the LOC device also has a support substrate, wherein the light sensor is an array of photodiodes on the support substrate that are registered with the probes. GAS123. Preferably, the photodiode array is less than 1,600 microns from the probe -143 - 201211533. GAS123. Preferably, the LOC device also has a CMOS circuit on the support substrate, the array of photodiodes being a component of the CMOS circuit, wherein when in use, the CMOS circuit responds to the detection of the negative control probe The ECL is emitted to cause an error signal. GAS123. Preferably, the LOC device also has a hybridization chamber comprising the probes and a pair of electrodes. GAS123. Preferably, the LOC device also has: a flow path containing the target fluid; wherein the CMOS circuit is interposed between the hybridization chamber and the support substrate, and the flow path attracts the fluid by capillary action To each of these hybrid chambers. GAS 123. Preferably, the LOC device also has a lysis unit, wherein the flow system contains a biological sample of the cell and the lysate destroys the cell membrane to release any genetic material therein. GAS123. Preferably, the hybridization chambers each have a wall portion that is transparent to the light emitted by the probe. GAS123. Il Preferably, the CMOS circuit is provided with the light sensor, wherein the wall portion is located between the probes and the light sensor. GAS 123.  Preferably, the wall portion is doped with a layer of cerium oxide. GAS 123. Preferably, the probe has a stem-loop structure, the loop portion of which contains a sequence complementary to the target nucleic acid sequence, the loop portion being located in the functional portion for quenching photon emission from the ECL luminophore Between the ECL luminophores, hybridization with the target nucleic acid sequence opens the loop portion and moves the ECL luminophore away from the functional portion. -144- 201211533 GAS123. Preferably, the CMOS circuit is configured to provide electrical pulses to the electrodes. GAS123. 15 Preferably, the electrical pulse has a value less than 0. During the 69 seconds period. GAS123. Preferably, the current of the electrical pulse is between 0. 1 nai to 69. 0 nai pei. GAS123. Preferably, the electrodes have an anode and a cathode, each having a finger-like configuration, the finger structures being configured to intersect the fingers of the anode with the fingers of the cathode. GAS123. 18 Preferably, the anode and cathode are separated by 0. 4 micron to 2 micron wide dielectric gap" GAS123. Preferably, the LOC device also has a PCR portion for amplifying the target nucleic acid sequence prior to detection by the probes. GAS 1 23. Preferably, the LOC device also has an upper cover, the upper cover being provided with a reagent reservoir for adding to the sample φ before detecting the target sequence, wherein the electrodes and the probes are interposed between the cover and Between these CMOS circuits. The hybridization array provides analysis of the target via hybridization and utilizes the control probe to improve the reliability of the analytical results. The electrochemiluminescence-based detection analysis eliminates the need for detection systems that require excitation light sources, excitation optics, and optical filter elements to provide a lighter and less expensive inspection system. It is not necessary to reject any excitation light and simplify the detector circuit, making the detection system even cheaper. GAS124. 1 This aspect of the invention provides a wafer-on-lab (LOC) device for detecting a nucleic acid sequence in a fluid of -145 to 201211533, the LOC device comprising: electrochemiluminescence for detecting the target nucleic acid sequence (ECL) a probe, each of which has an ECL luminophore for emitting photons in an excited state, a functional portion for quenching photon emission from the ECL luminophore by resonance energy transfer: for receiving electrical pulses An electrode for exciting the ECL luminophore; a hybridization chamber containing a probe for detecting the target and a pair of electrodes; and at least one positive control chamber containing a positive control probe having an ECL luminophore But there is no functional part for quenching photon emission. GAS124. Preferably, the LOC device also has a light sensor located adjacent to the probe for sensing those probes that respond to electrical pulses to produce ECL photons. GAS124. Preferably, the LOC device also has a support substrate, wherein the light sensor is a charge coupled device (CCD) array between the probe and the support substrate. GAS124. Preferably, the LOC device also has a support substrate, wherein the light sensor is an array of photodiodes on the support substrate that are registered with the hybrid chambers. GAS 124. Preferably, the photodiode array is less than 1,600 microns from the probe. GAS124. Preferably, the LOC device also has a CMOS circuit on the substrate supporting the -146-201211533, the array of photodiodes being a component of the CMOS circuit, wherein when used, the CMOS circuit response cannot be sensed from The ECL of the positive control probe emits an error signal. GAS124. Preferably, the LOC device also has at least one negative control chamber containing a negative control probe that is incapable of hybridizing to any of the nucleic acid sequences in the fluid. GAS124. Preferably, the LOC device also has: a flow path containing the target fluid; wherein the CMOS circuit is interposed between the hybridization chamber and the support substrate, and the flow path attracts the fluid by capillary action To each of these hybrid chambers" GAS124. Preferably, the LOC device also has a lysis unit, wherein the flow system contains a biological sample of the cell and the lysate destroys the cell membrane to release any genetic material therein. GAS124. Preferably, the hybridization chambers each have a wall portion that is transparent to the light emitted by the probe. GAS124. Il Preferably, the CMOS circuit is provided with the light sensor, wherein the wall portion is located between the probes and the light sensor. GAS 124. Preferably, the wall portion is doped with a layer of cerium oxide. GAS124. Preferably, the probe has a stem-loop structure, the loop portion of which contains a sequence complementary to the target nucleic acid sequence, the loop portion being located in the functional portion for quenching photon emission from the ECL luminophore Between the ECL luminophores, hybridization with the target nucleic acid sequence opens the loop portion and moves the ECL luminophore away from the functional portion. GAS124. Preferably, the C Μ Ο S circuit is configured to provide -147 - 201211533 electrical pulses to the electrodes. GAS 1 24. 1 5 Preferably, the electrical pulse has a value less than 0. During the 69 seconds period. GAS 1 24. 1 6 Preferably, the current of the electrical pulse is between 0. 1 nai to 69. 0 nai pei. GAS 1 24. Preferably, the electrodes have an anode and a cathode, each having a finger configuration, the finger structures being configured to intersect the fingers of the anode with the fingers of the cathode. GAS 124. 18 Preferably, the anode and cathode are separated by 0. Dielectric gap of 4 microns to 2 microns wide. GAS124. Preferably, the LOC device also has a PCR portion for amplifying the target nucleic acid sequence prior to detection by the probes. GAS 1 24. Preferably, the LOC device also has an upper cover, the upper cover being provided with a reagent reservoir for adding to the sample before detecting the target sequence, wherein the electrodes and the probes are interposed between the upper cover and the Between CMOS circuits. The hybridization array provides analysis of the target via hybridization and utilizes the control probe to improve the reliability of the analytical results. The electrochemiluminescence-based detection analysis eliminates the need for detection systems that require excitation light sources, excitation optics, and optical filter elements to provide a lighter and less expensive inspection system. It is not necessary to reject any excitation light and simplify the detector circuit, making the detection system even cheaper. GAS125. 1 This aspect of the invention provides a wafer-on-lab (LOC) device for detecting a target nucleic acid sequence in a fluid, the LOC loading -148 - 8 201211533 comprising: electrochemiluminescence for detecting the target nucleic acid sequence (ECL) probes, each of which has an ECL luminophore for emitting photons in an excited state, a functional moiety for quenching photon emission from the ECL luminophore by resonance energy transfer; An electrical pulse to excite the electrodes of the ECL luminophores; a hybridization chamber containing a probe for detecting the label and a pair of electrodes; and at least one negative control chamber containing a negative control probe that is incapable of Hybridization with any nucleic acid sequence in the fluid. GAS125. Preferably, the LOC device also has a light sensor located adjacent to the probe for sensing those probes that respond to electrical pulses to produce ECL photons. GAS125. Preferably, the LOC device also has a support substrate, wherein the light sensor is a charge φ coupling device (CCD) array between the probe and the support substrate. GAS125. Preferably, the LOC device also has a support substrate, wherein the light sensor is an array of photodiodes on the support substrate that are registered with the hybrid chambers. GAS125. Preferably, the photodiode array is less than 1,600 microns from the probe. GAS125. Preferably, the LOC device also has a CMOS circuit on the support substrate, the array of photodiodes being a component of the CMOS circuit, wherein when used, the CMOS circuit responds to detect from -149-201211533 The ECL of the negative control chamber emits an error signal. GAS125. Preferably, the LOC device also has at least one positive control chamber containing a positive control probe having an ECL luminophore but without a functional moiety for quenching photon emission. GAS 125. Preferably, the LOC device also has: a flow path containing the target fluid; wherein the CMOS circuit is interposed between the hybridization chamber and the support substrate, and the flow path attracts the fluid by capillary action To each of these hybrid chambers. GAS 125. Preferably, the LOC device also has a lysis unit, wherein the flow system contains a biological sample of the cell and the lysate destroys the cell membrane to release any genetic material therein. GAS125. Preferably, the hybridization chambers each have a wall portion that is transparent to the light emitted by the probe. GAS125. Il Preferably, the CMOS circuit is provided with the light sensor, wherein the wall portion is located between the probes and the light sensor. GAS125. Preferably, the wall portion is doped with a layer of cerium oxide. GAS125. Preferably, the probe has a stem-loop structure, the loop portion of which contains a sequence complementary to the target nucleic acid sequence, the loop portion being located in the functional portion for quenching photon emission from the ECL luminophore Between the ECL luminophores, hybridization with the target nucleic acid sequence opens the loop portion and moves the ECL luminophore away from the functional portion. GAS125. Preferably, the CMOS circuit is configured to provide electrical pulses to the electrodes. GAS125. 15 Preferably, the electrical pulse has a value less than 0. 69 seconds -150- 201211533 period. GAS125. Preferably, the current of the electrical pulse is between 0. 1 nai to 69. 0 nai pei. GAS 125. Preferably, the electrodes have an anode and a cathode, each having a finger configuration, the finger structures being configured to intersect the fingers of the anode with the fingers of the cathode. GAS125. 18 Preferably, the anode and cathode are separated by 0. 4 Between φ micron and 2 micron wide dielectric gap. GAS 1 25. Preferably, the LOC device also has a PCR portion for amplifying the target nucleic acid sequence prior to detection by the probes. GAS 1 25. Preferably, the LOC device also has an upper cover, the upper cover being provided with a reagent reservoir for adding to the sample before detecting the target sequence, wherein the electrodes and the probes are interposed between the upper cover and the Between CMOS circuits. The hybridization array provides analysis of the target via hybridization and utilizes the φ control probe to improve the reliability of the analytical results. The electrochemiluminescence-based detection analysis eliminates the need for detection systems that require excitation light sources, excitation optics, and optical filter elements to provide a lighter and less expensive inspection system. It is not necessary to reject any excitation light and simplify the detector circuit, making the detection system even cheaper. GAS 1 26. 1 This aspect of the invention provides a wafer-on-lab (LOC) device for detecting a target nucleic acid sequence in a fluid, the LOC device comprising: an electrochemiluminescent (ECL) probe for detecting the target nucleic acid sequence, -151 - 201211533 each of the probes has an ECL luminophore for emitting photons in an excited state, a functional portion for quenching photon emission from the ECL luminophore by resonance energy transfer; for receiving electrical pulses An electrode for exciting the ECL luminophore; a hybridization chamber containing a probe for detecting the label and a pair of electrodes; and at least one negative control chamber containing a negative control probe having no ECL luminophore. GAS126. Preferably, the LOC device also has a light sensor located adjacent to the probe for sensing those probes that respond to electrical pulses to produce ECL photons. GAS126. Preferably, the LOC device also has a support substrate, wherein the light sensor is a charge coupled device (CCD) array between the probe and the support substrate. GAS126. Preferably, the LOC device also has a support substrate, wherein the light sensor is an array of photodiodes on the support substrate that are registered with the hybrid chambers. GAS126. Preferably, the photodiode array is less than 1,600 microns from the probe. GAS126. Preferably, the LOC device also has a CMOS circuit on the support substrate, the array of photodiodes being a component of the CMOS circuit, wherein when used, the CMOS circuit responds to the detection from the negative control room. The ECL emits an error signal. GAS 126. Preferably, the LOC device also has at least one -152-201211533 positive control chamber containing a positive control probe having an ECL luminophore but without a functional moiety for quenching photon emission. GAS126. Preferably, the LOC device also has: a flow path containing the target fluid; wherein the CMOS' circuit is interposed between the hybridization chamber and the support substrate, and the flow path attracts the capillary by capillary action Fluid to each of the hybridization chambers. GAS126. Preferably, the LOC device also has a lysis unit, φ wherein the flow system contains a biological sample of the cell and the lysate destroys the cell membrane to release any genetic material therein. GAS126. Preferably, the hybridization chambers each have a wall portion that is transparent to the light emitted by the probe. GAS126. Il Preferably, the CMOS circuit is provided with the light sensor, wherein the wall portion is located between the probes and the light sensor. GAS126. Preferably, the wall portion is doped with a layer of cerium oxide. GAS126. Preferably, the probe has a stem-loop structure, the loop portion φ of which contains a sequence complementary to the target nucleic acid sequence, the loop portion being located in the functional portion for quenching photon emission from the ECL luminophore Between the ECL luminophore and the nucleic acid sequence of the target, the loop portion is opened and the ECL luminophore is moved away from the functional portion. GAS 126. Preferably, the CMOS circuit is configured to provide electrical pulses to the electrodes. GAS126. 15 Preferably, the electrical pulse has a value less than 0. During the 69 seconds period. GAS126. Preferably, the current of the electrical pulse is between 0. 1奈 -153- 201211533 amps to 69. 0 nai pei. GAS 1 26. Preferably, the electrodes have an anode and a cathode, each having a finger configuration, the finger structures being configured to intersect the fingers of the anode with the fingers of the cathode. GAS 1 26. 1 8 Preferably, the anode and the cathode are separated by 0. Dielectric gap of 4 microns to 2 microns wide. GAS126. Preferably, the LOC device also has a PCR portion for amplifying the target nucleic acid sequence prior to detection by the probes. GAS 1 26. Preferably, the LOC device also has an upper cover, the upper cover being provided with a reagent reservoir for adding to the sample before detecting the target sequence, wherein the electrodes and the probes are interposed between the upper cover and the C Μ Ο between S circuits. The hybridization array provides analysis of the target via hybridization and utilizes the control probe to improve the reliability of the analytical results. The electrochemiluminescence-based detection analysis eliminates the need for detection systems that require excitation light sources, excitation optics, and optical filter elements to provide a lighter and less expensive inspection system. It is not necessary to reject any excitation light and simplify the detector circuit, making the detection system even cheaper. GAS 127.  1 This aspect of the invention provides a wafer-on-lab (LOC) device for detecting a target nucleic acid sequence in a fluid, the LOC device comprising: an electrochemiluminescent (ECL) probe for detecting the target nucleic acid sequence, Each of the probes has an ECL luminophore for emitting photons in an excited state, and a functional portion for quenching photon emission from the ECL luminophore by resonance energy transfer: for receiving electricity Pulses to excite the electrodes of the ECL luminophores; a hybridization chamber containing a probe for detecting the label and a pair of electrodes: and at least one negative control chamber without the ECL probe. GAS127. Preferably, the LOC device also has a light sensor located adjacent to the probe for sensing those probes that respond to the electrical pulse φ to produce ECL photons. GAS127. Preferably, the LOC device also has a support substrate, wherein the light sensor is a charge coupled device (CCD) array between the probe and the support substrate. GAS127. Preferably, the LOC device also has a support substrate, wherein the light sensor is an array of photodiodes on the support substrate that are registered with the hybrid chambers. GAS127. Preferably, the photodiode array is less than 1,600 microns from the probe φ. GAS127. Preferably, the LOC device also has a CMOS circuit on the support substrate, the array of photodiodes being a component of the CMOS circuit, wherein when used, the CMOS circuit responds to the detection from the negative control room. The ECL emits an error signal. GAS127. Preferably, the LOC device also has at least one positive control chamber containing a positive control probe having an ECL luminophore but without a functional moiety for quenching photon emission. GAS127. Preferably, the LOC device also has: -155-201211533 a flow path containing the target fluid; wherein the CMOS circuit is interposed between the hybridization chamber and the support substrate, and the flow path is capillary The action draws the fluid to each of the hybrid chambers. GAS127. Preferably, the LOC device also has a lysis unit, wherein the flow system contains a biological sample of the cell and the lysate destroys the cell membrane to release any genetic material therein. GAS127. Preferably, the hybridization chambers each have a wall portion that is transparent to the light emitted by the probe. GAS127. Il Preferably, the CMOS circuit is provided with the light sensor, wherein the wall portion is located between the probes and the light sensor. GAS127. Preferably, the wall portion is doped with a layer of cerium oxide. GAS 127. Preferably, the probe has a stem-loop structure, the loop portion of which contains a sequence complementary to the target nucleic acid sequence, the loop portion being located in the functional portion for quenching photon emission from the ECL luminophore The ECL emits light between the oximes such that hybridization with the target nucleic acid sequence opens the loop portion and leaves the ECL luminophore away from the functional portion. GAS 12 7. Preferably, the CMOS circuit is configured to provide electrical pulses to the electrodes. GAS127. 15 Preferably, the electrical pulse has a value less than 0. During the 69 seconds period. GAS 12 7. 16 Preferably, the current of the electrical pulse is between 〇.  1 nai to 69. 0 nai pei. GAS 127. Preferably, the electrodes have an anode and a cathode, each having a finger configuration, the finger structures being configured to intersect the fingers of the anode - 156 - 201211533 with the fingers of the cathode. GAS127. 18 Preferably, the anode and cathode are separated by 0. Dielectric gap of 4 microns to 2 microns wide. GAS127. Preferably, the LOC device also has a PCR portion for amplifying the target nucleic acid sequence prior to detection by the probes. GAS 1 27. Preferably, the LOC device also has an upper cover, the upper cover being provided with a reagent reservoir for adding to the sample φ before detecting the target sequence, wherein the electrodes and the probes are interposed between the cover and Between the C Μ Ο S circuits. The hybridization array provides analysis of the target via hybridization and utilizes the control probe to improve the reliability of the analytical results. The electrochemiluminescence-based detection analysis eliminates the need for detection systems that require excitation light sources, excitation optics, and optical filter elements to provide a lighter and less expensive inspection system. It is not necessary to reject any excitation light and simplify the detector circuit, making the detection system even cheaper. φ GAS128. 1 This aspect of the invention provides a wafer-on-lab (LOC) device for detecting a target nucleic acid sequence in a fluid, the LOC device comprising: an electrochemiluminescent (ECL) probe for detecting the target nucleic acid sequence, Each of the probes has an ECL luminophore for emitting photons in an excited state, a functional portion for quenching photon emission from the ECL luminophore by resonance energy transfer; for receiving electrical pulses to excite such Electrode of an ECL luminophore; detection for exposure to photons emitted by the ECL luminophore - 157 - 201211533 Diode: and at least one calibrated photodiode for exposure to ambient light; The difference between the output from any of the sense photodiodes and the output from the calibrated photodiode is compared to a predetermined valve 値 difference, the output difference being greater than the predetermined valve 値 indicating the presence of the target system. GAS128. Preferably, the LOC device further comprises: a hybridization chamber containing a probe for detecting the target and a pair of electrodes, wherein the detection photodiode system is respectively located in registration with each of the hybrid chambers: and is located adjacent to the calibration photoelectric The calibration chamber for the diode. GAS 128. Preferably, the LOC device also has a plurality of calibration chambers and a plurality of corresponding calibrated photodiodes distributed throughout the array of hybridization chambers, wherein, when used, from the detection photodiodes The output of one is compared to the output from the calibrated photodiode closest to the detection photodiode. GAS128. Preferably, each of the calibration chambers is surrounded by a three by three hybrid chamber block. GAS128. Preferably, the distance between the detection photodiode and the hybridization chamber is less than 1,600 microns. GAS 128. Preferably, the LOC device also has a CMOS circuit on the support substrate, the detection and calibration of the components of the CMOS circuit of the photodiode system, wherein when used, the CMOS circuit has a function for measuring each of the detection electrodes Comparator circuit for the difference in output between the diode and the closest calibrated photodiode - 158 - 201211533 GAS128. Preferably, the calibration chamber has a probe that lacks the ECL radiant group but includes the functional portion for quenching photon emission. GAS 1 28. Preferably, the LOC device also has: a flow path containing the target fluid; wherein the CMOS circuit is interposed between the hybridization chamber and the support substrate, and the flow path attracts the fluid by capillary action To each of these hybrid chambers. GAS128. Preferably, the LOC device also has a lysis unit, φ wherein the flow system contains a biological sample of the cell and the lysate destroys the cell membrane to release any genetic material therein. GAS128. Preferably, the hybridization chambers each have a wall portion that is transparent to the light emitted by the probe. GAS128. Il Preferably, the CMOS circuit is provided with the light sensor, wherein the wall portion is located between the probes and the light sensor. GAS128. Preferably, the wall portion is doped with a layer of cerium oxide. GAS128. Preferably, the probe has a stem-loop structure, the loop portion φ of which contains a sequence complementary to the target nucleic acid sequence, the loop portion being located in the functional portion for quenching photon emission from the ECL luminophore Between the ECL luminophore and the nucleic acid sequence of the target, the loop portion is opened and the ECL luminophore is moved away from the functional portion. GAS 128. Preferably, the CMOS circuit is configured to provide electrical pulses to the electrodes. GAS128. 15 Preferably, the electrical pulse has a value less than 0. During the 69 seconds period. GAS128. Preferably, the current of the electrical pulse is between 0. 1奈 -159- 201211533 amp to 69. 0 nai pei. GAS128. Preferably, the electrodes have separate anodes and cathodes, the fingers being configured to intersect the fingers of the cathode. GAS128. Preferably, a dielectric gap of between microns and 2 microns is provided between the anode and the cathode. GAS128. Preferably, the LOC device is also narrowly used to amplify the target core prior to detection by the probes. [ GAS 1 28. Preferably, the LOC device is also provided with a reagent reservoir for detecting the sequence of the sample, wherein the electrodes and the probes are interposed between the circuits of the C Μ Ο S. The hybrid array provides analysis of the target via hybridization. The calibration light sensor improves the reliability and sensitivity of the results. The electrochemiluminescence-based detection and analysis system is a lighter and cheaper detection system for detection systems that are free of light sources, excitation optics, and optical filter elements. It is not necessary to disapprove the detector circuit, making the detection system even cheaper. GAS 129. 1 This aspect of the invention provides a wafer-on-lab (LOC) device for use in a nucleic acid sequence in use, comprising: a hybridization chamber comprising an electrochemical hairpin for detecting the target nucleic acid sequence, each of the probes Having a finger shape in an excited state to isolate the anode system. 4 ί PCR section, 唆 sequence. There is an upper cover, which is added to the upper cover and the above, and utilizes the property and the dynamic range to eliminate the need for excitation, thereby providing excitation light and simply detecting the fluid. The LOC light-emitting (ECL) probe emits photons - 160 - 201211533 EC L luminophore, a functional moiety for quenching photon emission from the EC L luminophore by resonance energy transfer; at least one calibration chamber comprising an ECL probe designed to be complementary to any nucleic acid sequence in the fluid And electrodes for receiving electrical pulses to excite the ECL luminophores. GAS 129. 2 Preferably, the LOC device also has: a detecting photo φ diode for exposing photons emitted by the ECL illuminating group; and at least one calibrated photodiode for exposing to ambient light; wherein In use, the difference between the output from any of the sense photodiodes and the output from the calibrated photodiode is compared to a predetermined valve , difference that is greater than the predetermined valve 値 indicating the presence of the target. GAS129. Preferably, the LOC device also has a plurality of calibration chambers and a plurality of corresponding calibrated photodiode φ pole bodies distributed throughout the array of hybridization chambers, wherein when used, from the detection photodiode The output of either is compared to the output from the calibrated photodiode closest to the detection photodiode. GAS129. Preferably, each of the calibration chambers is surrounded by a three by three hybrid chamber block. GAS129. Preferably, the distance between the detection photodiode and the hybridization chamber is less than 1,600 microns. GAS129. Preferably, the LOC device also has a CMOS circuit on the support substrate, the detection and calibration of the photodiode system of the -161 - 201211533 CMOS circuit component, wherein when used, the CMOS circuit has a The comparator circuit detects the difference in output between the photodiode and the closest calibrated photodiode. GAS129. Preferably, the calibration chamber has a probe that lacks the ECL radiant group but includes the functional portion for quenching photon emission. GAS129. Preferably, the LOC device also has: a flow path containing the target fluid; wherein the CMOS circuit is interposed between the hybridization chamber and the support substrate, and the flow path attracts the fluid by capillary action To each of these hybrid chambers. GAS129. Preferably, the LOC device also has a lysis unit, wherein the flow system contains a biological sample of the cell and the lysate destroys the cell membrane to release any genetic material therein. ' GAS129. Preferably, the hybridization chambers each have a wall portion that is transparent to the light emitted by the probe. GAS129. Il Preferably, the CMOS circuit is provided with the light sensor, wherein the wall portion is located between the probes and the light sensor. GAS 129. Preferably, the wall portion is doped with a layer of cerium oxide. GAS 129. Preferably, the probe has a stem-loop structure, the loop portion of which contains a sequence complementary to the target nucleic acid sequence, the loop portion being located in the functional portion for quenching photon emission from the ECL luminophore Between the ECL luminophores, hybridization with the target nucleic acid sequence opens the loop portion and moves the ECL luminophore away from the functional portion. GAS 1 29. Preferably, the C Μ Ο S circuit is configured to provide electrical pulses to the electrodes. -162- 201211533 GAS129. 15 Preferably, the electrical pulse has a value less than 0. During the 69 seconds period. GAS129. 16 Preferably, the current of the electrical pulse is between 〇. 1 nai to 69. 0 nai pei. GAS129. Preferably, the electrodes have an anode and a cathode, each having a finger configuration, the finger structures being configured to intersect the fingers of the anode with the fingers of the cathode. GAS 129. 18 Preferably, the anode and cathode are separated by 0. Dielectric gap of 4 microns to 2 microns wide. GAS 1 29. Preferably, the LOC device also has a PCR portion for amplifying the target nucleic acid sequence prior to detection by the probes. GAS 1 29. Preferably, the LOC device also has an upper cover, the upper cover being provided with a reagent reservoir for adding to the sample before detecting the target sequence, wherein the electrodes and the probes are interposed between the upper cover and the C Μ Ο between S circuits. The hybridization array provides analysis of the target via hybridization and utilizes the calibration probe to improve the reliability, sensitivity, and dynamic range of the analytical results. The electrochemiluminescence-based detection analysis eliminates the need for detection systems that require excitation light sources, excitation optics, and optical filter elements to provide a lighter and less expensive inspection system. It is not necessary to reject any excitation light and simplify the detector circuit, making the detection system even cheaper. GAS 1 3 0 · 1 This aspect of the invention provides a wafer-on-lab (LOC) device for detecting a target nucleic acid sequence in a fluid, the LOC device comprising: -163-201211533 for detecting the target nucleic acid sequence Electrochemiluminescence (EC L) probes, each of which has an Ecl luminophore for emitting photons in an excited state, and a function for quenching photon emission from the ECL luminophore by resonance energy transfer a portion; a calibration probe that does not have an ECL luminophore; and an electrode that receives an electrical pulse to excite the ECL luminophore. GAS130. 2 Preferably, the LOC device also has: a detection photodiode for exposing photons emitted by the ECL luminophore; and at least one calibration photodiode for exposing to the calibration probe; When in use, the difference between the output from any of the sense photodiodes and the output from the calibrated photodiode is compared to a predetermined valve 値 difference that is greater than the predetermined valve 値 indicating the presence of the target. GAS130. Preferably, the LOC device also has an array of calibration chambers for the ECL probes and a plurality of calibration chambers distributed throughout the array of hybridization chambers, each of the calibration chambers including the calibration probes and each The calibration chambers have corresponding calibration photodiodes, wherein when used, the output from either of the detection photodiodes and the output from the calibration photodiode closest to the detection photodiode Comparison. GAS130. Preferably, each of the calibration chambers is surrounded by a three by three hybrid chamber block. GAS 130. Preferably, the distance between the detection photodiode and the hybridization chamber is less than 1,600 microns. -164- 201211533 GAS130. Preferably, the LOC device also has a CMOS circuit on the substrate, the component for detecting and calibrating the photodiode CMOS circuit, wherein when used, the CMOS circuit is configured to measure each of the detecting photodiodes A comparator circuit that differs from the output of the closest calibration photocell. GAS 13 0. Preferably, the calibration probe includes a functional portion for quenching the emission. φ GAS130. Preferably, the LOC device also has: a flow path containing the target fluid; wherein the CMOS circuit is interposed between the hybridization chamber and the support substrate, and the flow path attracts the fluid to each by capillary action These hybrid chambers. GAS130. Preferably, the LOC device also has a lysate wherein the flow system contains a biological sample of cells and the lysate destroys the membrane to release any genetic material therein. GAS130. Preferably, the hybridization chambers each have a wall portion that is transparent to the light emitted by the φ. GAS130. Preferably, the CMOS circuit has the photo-electric body, wherein the wall portion is located between the probe and the photodiode. 12 Preferably, the wall is entangled with GAS 130 of cerium oxide. Preferably, the ECL probe has a stem loop junction whose loop portion contains a sequence complementary to the target nucleic acid sequence, the loop portion being located in the function for quenching photon emission from the ECL luminophore and the ECL luminescence Between the clusters, the loop portion is hybridized to the target nucleic acid sequence and the ECL luminophore is moved away from the functional portion. Support for this useful two-pole photon, part, and cell probe bipolar 〇 layer. Structure, Division Department Open -165- 201211533 GAS 13 0. Preferably, the CMOS circuit is configured to provide electrical pulses to the electrodes. GAS130. 15 Preferably, the electrical pulse has a value less than 0. During the 69 seconds period. GAS130. Preferably, the current of the electrical pulse is between 0. 1 nai to 69. 0 nai pei. GAS130. Preferably, the electrodes have an anode and a cathode, each having a finger configuration, the finger structures being configured to intersect the fingers of the anode with the fingers of the cathode. GAS130. 18 Preferably, the anode and cathode are separated by 0. Dielectric gap of 4 microns to 2 microns wide. GAS 1 30. Preferably, the LOC device also has a PCR portion for amplifying the target nucleic acid sequence prior to detection by the probes. GAS 1 30. Preferably, the LOC device also has an upper cover, the upper cover being provided with a reagent reservoir for adding to the sample before detecting the target sequence, wherein the electrodes and the probes are interposed between the upper cover and the C Μ Ο between S circuits. The hybridization array provides analysis of the target via hybridization and utilizes the calibration probe to improve the reliability, sensitivity, and dynamic range of the analytical results. The electrochemiluminescence-based detection analysis eliminates the need for detection systems that require excitation light sources, excitation optics, and optical filter elements to provide a lighter and less expensive inspection system. It is not necessary to reject any excitation light and simplify the detector circuit, making the detection system even cheaper. GAS131. 1 This aspect of the invention provides a wafer-on-lab (LOC) device for detecting a nucleic acid sequence of a target in a fluid of -166 to 201211533, the LOC device comprising: electrochemiluminescence comprising a nucleic acid sequence for detecting the target ( ECL) hybridization chambers for probes, each of the ECL probes having an ECL luminophore for emitting photons in an excited state, and a functional portion for quenching photon emission from the ECL luminophore by resonance energy transfer; At least one calibration φ chamber comprising a probe sealed to be in contact with the fluid; and an electrode for receiving an electrical pulse to excite the ECL luminophores. GAS131. 2 Preferably, the LOC device also has: a detection photodiode for exposing photons emitted by the ECL luminophore; and at least one calibration photodiode for exposing to the calibration chamber; In use, the difference between the output from any of the sense photodiodes and the output from the calibrated photo φ diode is compared to a predetermined valve , difference that is greater than the predetermined valve 値 indicating the presence of the target. GAS131. Preferably, the LOC device also has a plurality of calibration chambers distributed throughout the array of hybridization chambers, each of the calibration chambers having a corresponding calibration photodiode, wherein when used, the detection photodiode The output of any of the bodies is compared to the output from the calibrated photodiode closest to the detection photodiode. GAS13 1. Preferably, each of the calibration chambers is surrounded by a three by three hybrid chamber block. 167 201211533 GAS131. Preferably, the distance between the detection photodiode and the hybridization chamber is less than 1,600 microns. GAS131. Preferably, the LOC device also has a CMOS circuit on the support substrate, the detection and calibration of the components of the CMOS circuit of the photodiode system, wherein when used, the CMOS circuit has a function for measuring each of the detection electrodes A comparator circuit for the difference in output between the diode and the closest calibrated photodiode. GAS131. Preferably, the LOC device also has: a flow path containing the target fluids, wherein the hybridization chambers each have an inlet for fluid communication between the flow path and the probes, and the at least one calibration The chamber is separated from the flow path. GAS131. Preferably, the CMOS circuit is interposed between the hybridization chambers and the support substrate, and the flow path attracts the fluid to each of the hybridization chambers by capillary action. GAS131. Preferably, the LOC device also has a lysis unit, wherein the flow system contains a biological sample of the cell and the lysate destroys the cell membrane to release any genetic material therein. GAS 13 1. Preferably, the hybridization chambers each have a wall portion that is transparent to the light emitted by the probe. GAS131. Preferably, the CMOS circuit is provided with the photodiode, wherein the wall portion is located between the probe and the photodiode. GAS 13 1. Preferably, the wall portion is doped with a layer of cerium oxide. GAS 13 1. Preferably, the ECL probe has a stem-loop structure, the loop portion of which contains a sequence complementary to the target nucleic acid sequence, the loop portion is -168-201211533 located for quenching photon emission from the EC L luminophore Between the functional portion and the ECL luminophore, such that hybridization with the target nucleic acid sequence opens the loop portion and moves the ECL luminophore away from the functional portion. GAS 13 1. Preferably, the CMOS circuit is configured to provide electrical pulses to the electrodes. GAS131. 15 Preferably, the electrical pulse has a value less than 0. During the 69 seconds period. Φ GAS131. Preferably, the current of the electrical pulse is between 0. 1 nai to 69. 0 nai pei. GAS 1 3 1 .  Preferably, the electrodes have an anode and a cathode, each having a finger configuration, the finger structures being configured to intersect the fingers of the anode with the fingers of the cathode. GAS131. 18 Preferably, the anode and cathode are separated by 0. Dielectric gap of 4 microns to 2 microns wide. GAS131. Preferably, the LOC device also has a PCR portion, φ which is used to amplify the target nucleic acid sequence prior to detection by the probes. GAS131. Preferably, the LOC device also has an upper cover, the upper cover being provided with a reagent reservoir for adding to the sample before detecting the target sequence, wherein the electrodes and the probes are interposed between the upper cover and the Between CMOS circuits. The hybridization array provides analysis of the target via hybridization and utilizes the calibration probe to improve the reliability, sensitivity, and dynamic range of the analytical results. The electrochemiluminescence-based detection analysis eliminates the need for detection systems that require excitation light sources, excitation optics, and optical filter elements to provide a lighter and less expensive inspection system. It is not necessary to reject any excitation light and simplify the detector circuit, making the detection system even cheaper. GAS 132. 1 This aspect of the invention provides a wafer-on-lab (LOC) device for detecting a target nucleic acid sequence in a fluid, the LOC device comprising: an electrochemiluminescent (ECL) probe for detecting the target nucleic acid sequence, Each of the ECL probes has an ECL luminophore for emitting photons in an excited state, a functional portion for quenching photon emission from the ECL luminophore by resonant energy transfer; at least one configured to generate a calibrated emission a calibration source; an electrode for receiving an electrical pulse to excite the ECL luminophore; at least one detection photodiode for sensing photon emission from the ECL luminophore and generating a detection output; at least one for sensing The calibration emits and produces a calibrated output of the calibrated photodiode; and a CMOS circuit having a differential circuit for subtracting the calibrated output from the sense output. GAS 1 32. Preferably, the LOC device also has a plurality of calibration sources and a plurality of corresponding calibration photodiodes, and a plurality of detection photodiodes respectively registered with the ECL probes. GAS132. Preferably, the calibration source is a calibration probe that does not have an ECL luminophore. GAS 1 32. Preferably, the LOC device also has an array of hybridization chambers containing the ECL probes and a plurality of calibration chambers containing calibration sources distributed throughout the array of hybridization chambers -170-201211533, wherein when used, from The output of any of the diodes is compared to the output of the calibrated photodiode from the closest detector. GAS132. Preferably, the calibration source calibration probe chamber is configured to seal the calibration probe from the nuclear fluid containing the target. GAS132. Preferably, each of the calibration chambers is surrounded by a three-square chamber block. GAS 132. Preferably, the detection photodiode is at a distance of less than 1,600 microns. GAS132. Preferably, the calibration probe includes a functional portion for transmission. GAS132. Preferably, the LOC device also has: a flow path containing the target fluid; wherein the CMOS circuit is interposed between the hybridization chamber and the support base φ and the flow path attracts the fluid to each by capillary action These hybrids: GAS132. Preferably, the LOC device also has a biological sample in which the flow system contains cells and the lysate is ruptured to release any genetic material therein. GAS132. Il Preferably, the hybridization chambers each have a wall portion through which the light emitted by the probe penetrates. GAS132. 12 Preferably, the wall is enthalpy of GAS 1 32. Preferably, the ECL probe has a loop portion of the stem containing a sequence complementary to the target nucleic acid sequence, the detection photodiode and the third of the calibration acid sequence hybridize between the hybridization chamber quenching photonic plates, In the lysis part, the cell is broken by the layer of the ECL. a ring structure, a loop moiety -171 - 201211533, located between the functional moiety for quenching photon emission from the ECL luminophore and the ECL luminophore such that hybridization with the target nucleic acid sequence opens the loop portion and The ECL luminophore is remote from the functional portion. GAS 13 2. Preferably, the CMOS circuit is configured to provide electrical pulses to the electrodes. GAS132. 15 Preferably, the electrical pulse has a value less than 0. During the 69 seconds period. GAS132. Preferably, the current of the electrical pulse is between 0. 1 nai to 69. 0 nai pei. GAS 13 2. Preferably, the electrodes have an anode and a cathode, each having a finger configuration, the finger structures being configured to intersect the fingers of the anode with the fingers of the cathode. GAS132. 18 Preferably, the anode and cathode are separated by 0. Dielectric gap of 4 microns to 2 microns wide. GAS 13 2. Preferably, the LOC device also has a PCR portion for amplifying the target nucleic acid sequence prior to detection by the probes. GAS 1 3 2. Preferably, the LOC device also has an upper cover, the upper cover being provided with a reagent reservoir for adding to the fluid prior to detecting the sequence of the target, wherein the electrodes and the probes are interposed between the upper cover and the Between CMOS circuits. The hybrid array provides analysis of the target via hybridization and utilizes the calibration circuit to improve the reliability, sensitivity, and dynamic range of the analytical results. The detection and analysis of the target based on electrochemiluminescence eliminates the need for a detection system that requires an excitation source, an excitation optical, and an optical filter element, thereby providing -172-

201211533 Lighter and cheaper detection system. There is no need to disapprove the detector circuit, making the detection system even cheaper. GAS 13 3.1 This aspect of the invention provides a microfluidic test module for the nucleic acid sequence of the target, which is configured for hand movement An outer casing having an inlet for a fluid of the target nucleic acid sequence; a hybridization chamber mounted in the outer casing, the electrochemiluminescent (ECL) probe of the nucleic acid sequence of the hybridization chamber label, each having an excitation state ECL luminescence resonance energy transfer of the emitted photon quenches the photon portion from the ECL luminescence group, and is used to receive an electrical pulse to excite the ECL luminescence. The volume of the hybridization chamber is less than 900,000 cubic microns GAS 133.2. Preferably, the hybridization chamber The volume is cubic micron. GAS 133.3 Preferably, the body of the hybridization chamber is 1 cubic micrometer. GAS 133.4 Preferably, the hybridization chamber is a micron in volume. GAS 133.5 Preferably, the microfluidic test mode is for exposure to a photodetector emitted by the ECL luminophore; and for providing electrical pulses to the electrodes for control. Circuit GAS 133.6 preferably, the micro The fluid test mode and the excitation light are also used for the detection of the fluid test module. The method includes: an electrode for receiving an ECL probe group for detecting the | and a functional group for emitting by the electron; the 〇E is smaller than 2 0 0,0 0 0 Really less than 40,000 Less than 9,0 0 The vertical group also has: The sub-detection optical sensation group also has: -173- 201211533 The communication interface of the control circuit transmits data to the external device. GAS 133.7 Preferably, the microfluidic test module also has an array of hybridization chambers containing ECL probes for different target nucleic acid sequences, wherein the control circuit has memory for storage Identification data for ECL probes in these hybrid chambers. GAS 133.8 Preferably, the communication interface is a universal serial bus (USB) connector such that the housing is configured as a USB flash drive. GAS 133.9 Preferably, the detection light sensor registers the detection photodiode array of the hybrid chambers. GAS 1 3 3 .1 0 Preferably, the microfluidic test module also has: at least one calibration source for providing a calibration emission, and a calibration light sensor for sensing the calibration emission, wherein the control circuit has A differential circuit is operative to subtract the calibrated light sensor output from the detected light sensor output. Preferably, the microfluidic test module also has a plurality of calibration sources, wherein the detection light sensor is a photodiode array respectively registered with each of the ECL probes, and the calibration light sensor A plurality of calibrated photodiodes respectively registered with the calibration sources. GAS 133.12 Preferably, the calibration source is a calibration probe that does not have an ECL luminophore. GAS 1 3 3 .1 3 Preferably, the microfluidic test module also has a plurality of calibration chambers, the calibration chamber containing calibration sources distributed throughout the array of hybridization chambers, wherein when used, the detection photodiodes are The output of any of the polar bodies is compared to the output from the calibrated photodiode closest to the detection photodiode. Preferably, the calibration source is a calibration probe and the calibration chamber is configured to seal the calibration probe from the fluid containing the target nucleic acid sequence. GAS 133.15 Preferably, each of the calibration chambers is surrounded by a three by three hybrid cell block. GAS 133.16 Preferably, the distance between the detection photodiode and the hybridization chamber is less than 1,600 microns. GAS 13 3.17 Preferably, the calibration probe includes a functional portion for quenching photon emission. GAS133.18 Preferably, the ECL probe has a stem-loop structure, the loop portion of which contains a sequence complementary to the target nucleic acid sequence, the loop portion being located in the functionality for quenching photon emission from the ECL luminophore Portion between the portion and the ECL luminophore to hybridize with the target nucleic acid sequence to open the loop portion and to move the ECL luminophore away from the functional portion. GAS 133.19 Preferably, the electrical pulse transmitted to the electrode is a DC pulse and has a period of less than 0.69 seconds. GAS 1 3 3.20 Preferably, the electrodes have an anode and a cathode, each having a finger configuration, the finger structures being configured to intersect the fingers of the anode with the fingers of the cathode. This low volume hybridization chamber represents, to some extent, a low probe volume, thereby providing low probe cost and an inexpensive detection system. The electrochemiluminescence-based detection analysis eliminates the need for a detection system that requires an excitation source, excitation optics, and optical filter elements to provide a lighter and less expensive detection system. There is no need to reject any excitation light. -175- 201211533 This detector circuit makes the detection system even cheaper. GAS 134.1 This aspect of the invention provides a microfluidic test module for detecting a target nucleic acid sequence in a fluid, the test module comprising: an outer casing having an inlet for receiving a nucleic acid sequence containing the target a fluid pair; an electrode pair for receiving an electrical pulse; respectively spotting an electrochemiluminescence (ECL) probe in contact with each of the pair of electrodes, the ECL probe spotting comprising an ECL having a photon for emitting in an excited state a luminophore and an ECL probe for quenching a functional portion of photon emission from the ECL luminophore by resonance energy transfer such that an electrical pulse transmitted to the pair of electrodes excites the ECL luminophore; The ECL probe mass in the probe spot is less than 270 picograms. Preferably, GAS 1 34.2 is less than 60 picograms. Preferably, GAS 1 34.3 is less than 12 picograms. Preferably, GAS 1 34.4 is less than 2.7 picograms. GAS 1 34.5 Preferably, a ligand complex. The probe mass in each probe spot is the probe mass in each probe spot. The probe mass in each probe spot has the transition metal _ GAS 134.6. The microfluidic test module also has: a detection light sensor for exposing to photons emitted by the ECL luminophore; and a control circuit for supplying electrical pulses to the electrodes. -176- 201211533 GAS 134.7 Preferably, the microfluidic test module also has a communication interface of the control circuit for transmitting data to an external device. GAS 134.8 Preferably, the microfluidic test module also has an array of chambers containing probes for different target nucleic acid sequences, wherein the control circuit has memory for storage in an equal hybridization chamber Identification of ECL probes. GAS 134.9 Preferably, the hybridization chambers have a volume of 900,000 cubic microns. GAS 134.10 Preferably, the hybridization chambers have a volume of 200,000 cubic microns. GAS 134.il Preferably, the communication interface is a universal serial (USB) connector such that the housing is configured as a USB flash drive GAS 134.12. Preferably, the detection light sensor is registered with the The detection of the photodiode array of the chamber. GAS134.13 Preferably, the microfluidic test module also has at least one calibration source for providing a calibration emission and at least one calibration photodiode for sensing the calibration emission, wherein the control circuit has a differential circuit to For subtracting the photodiode output from each of the detected photodiode outputs. GAS134.14 Preferably, the microfluidic test module also has a calibration source and a plurality of corresponding calibration photodiodes, and the calibration photodiodes are respectively registered with the calibration source. GAS 1 34.1 5 Preferably, the calibration source is a calibration probe hybridization ECL that does not have an ECL, and each of the J, J, J, and the like is used to calibrate a plurality of polar body light groups -177 - 201211533 GAS134 .16 Preferably, the microfluidic test module also has a plurality of calibration chambers. The calibration chamber contains calibration sources distributed throughout the hybrid chambers, wherein when used, 'from the detection photodiode The output of one is compared to the output from the calibrated photodiode closest to the detection photodiode. GAS 134·17 Preferably, the calibration source is a calibration probe and the calibration chamber is configured to seal the calibration probe from the fluid containing the target nucleic acid sequence. GAS 134.18 Preferably, each of the calibration chambers is surrounded by a three by three hybrid cell block. GAS 134.19 Preferably, the distance between the detection photodiode and the hybridization chamber is less than 1,600 microns. GAS 1 34.20 Preferably, the ECL probe has a stem-loop structure, the loop portion of which contains a sequence complementary to the target nucleic acid sequence, the loop portion being located in the functionality for quenching photon emission from the ECL luminophore Portion between the portion and the ECL luminophore to hybridize with the target nucleic acid sequence to open the loop portion and to move the ECL luminophore away from the functional portion. This low probe volume represents a low probe cost, which in turn allows for this inexpensive detection system. The electrochemiluminescence-based detection analysis eliminates the need for detection systems that require excitation light sources, excitation optics, and optical filter elements to provide a lighter and less expensive inspection system. It is not necessary to reject any excitation light and simplify the detector circuit, making the detection system even cheaper. GAS 135.1 This aspect of the invention provides a test module for detecting a nucleic acid sequence of a target in a fluid of 178-201211533, the test module comprising: a housing having a container for receiving a nucleic acid containing the target a fluid of sequence; an electrochemiluminescence (ECL) probe having an ECL luminophore that emits photons when in an excited state and a functional moiety that quenches photon emission from the ECL luminophore by resonance energy transfer; Electrical pulses to excite the electrodes of the ECL luminophores; and detection light sensors for exposure to photons emitted by the ECL luminophores. GAS 1 35.2 Preferably, the test module also has a lab-on-a-chip (LOC) device, the LOC device being provided with circuitry for providing electrical pulses to the electrodes, wherein the detection light sensor is plugged into the circuit And the ECL probe, the electrode, and the control circuit are integrated into the LOC device. GAS 135.3 Preferably, the LOC device has a support substrate for supporting the circuit, which in turn supports the electrodes and ECL probes. GAS 1 35.4 Preferably, the circuit has a CMOS circuit layer deposited on the support substrate for providing electrical pulses to the electrodes. GAS 1 35.5 Preferably, the electrical pulse has a period of less than 0.69 seconds. GAS 1 35.6 Preferably, the electrical current of the electrical pulse is between 0.1 nanoamperes and 69.0 nanoamperes. GAS 1 35.7 Preferably, the test module also has: a communication interface of the circuit for transmitting data to an external device. GAS 1 35.8 Preferably, the test module also has a hybrid cell array -179-201211533, the hybridization chamber containing ECL probes for different target nucleic acid sequences, wherein the CMOS circuit has memory for storage Identification data for ECL probes in each of these hybrid chambers. Preferably, the GAS 135.9 has a volume of less than 900,000 cubic microns. GAS 135.10 Preferably, the hybridization chambers have a volume of less than 200,000 cubic microns. GAS 135.11 Preferably, the communication interface is a universal serial bus (USB) connector such that the housing is configured as a USB flash drive. GAS 1 3 5 . 1 2 Preferably, the detection light sensor registers the detection photodiode arrays of the hybrid chambers. GAS 1 3 5 · 1 3 Preferably, the test module also has at least one calibration source for providing a calibration emission, and at least one calibration photodiode for sensing the calibration emission, wherein the CMOS circuit has A differential circuit subtracts the calibrated photodiode output from the output of one or more of the detection photodiodes. GAS 1 3 5 · 1 4 Preferably, the test module also has a plurality of calibration sources and a plurality of corresponding calibration photodiodes, the calibration photodiodes being respectively registered with the calibration source. GAS 135.15 Preferably, the calibration source is a calibration probe that does not have an ECL luminophore. GAS 1 3 5 · 1 6 Preferably, the test module also has a plurality of calibration chambers. The calibration chamber contains calibration sources distributed throughout the array of hybridization chambers, wherein when used, the detection photodiodes are used. The output of either one is compared to the output of 180-201211533 from the calibrated photodiode closest to the detection photodiode. GAS 13 5.17 Preferably, the calibration source is a calibration probe and the calibration chamber is configured to seal the calibration probe from the fluid containing the target nucleic acid sequence. GAS 135.18 Preferably, each of the calibration chambers is surrounded by a three by three hybrid cell block. φ GAS 135.19 Preferably, the distance between the detection photodiode and the hybridization chamber is less than 1,600 microns. GAS 1 35.20 Preferably, the ECL probe has a stem-loop structure, the loop portion of which contains a sequence complementary to the target nucleic acid sequence, the loop portion being located in the functionality for quenching photon emission from the ECL luminophore Portion between the portion and the ECL luminophore to hybridize with the target nucleic acid sequence to open the loop portion and to move the ECL luminophore away from the functional portion. The integrated φ image sensor with driver for exciting the electroluminescent luminophore eliminates the need for expensive external imaging systems, providing a comprehensive solution that is mass-produced and inexpensive, with low system component counts representing light weight and height Mobility system. The integrated image sensor has the benefit of increased read sensitivity due to large angle light collection and eliminates the need to use optical components in the string of light collecting elements. The electrochemiluminescence-based detection analysis eliminates the need for detection systems that require excitation light sources, excitation optics, and optical filter elements to provide a lighter and less expensive inspection system. It is not necessary to reject any excitation light and simplify the detector circuit, making the detection system even cheaper. -181 - 201211533 GAS 13 6.1 Aspects of the invention provide a portable test module for exciting an electrochemiluminescent probe configured to detect a target nucleic acid sequence, the test module comprising: configured for hand movement a housing having a container for receiving a fluid containing the target nucleic acid sequence; an electrochemiluminescence (ECL) probe having an ECL luminophore that emits photons when excited, and quenching by resonance energy transfer a functional portion of photon emission from the ECL luminophore; and an electrode for receiving an electrical pulse to excite the ECL luminophore; wherein when used, the ECL probe of one of the labeled nucleic acid sequences has been detected The needle is reconfigured such that the functional portion does not quench photon emission from the ECL luminophore when excited by the electrodes. GAS 136.2 Preferably, the portable test module also has circuitry for providing the electrical pulse to the electrodes, the circuit having a detection light sensor for exposure to photons emitted by the ECL luminophore . GAS 136.3 Preferably, the portable test module also has a lab-on-a-chip (LOC) device having a support substrate for supporting the circuit, which in turn supports the electrodes and ECL probes. GAS 136.4 Preferably, the circuit has a CMOS circuit layer deposited on the support substrate for providing electrical pulses to the electrodes. GAS 136.5 Preferably, the electrical pulse has a period of less than 0.69 seconds. GAS 136.6 Preferably, the current of the electrical pulse is between 0.1 Na 8 -182 - 201211533 amps to 69.0 Nai amps. GAS 136.7 Preferably, the expandable test module also has: a communication interface of the circuit for transmitting data to an external device. GAS 136.8 Preferably, the portable test module also has an array of hybridization chambers containing ECL probes for different target nucleic acid sequences 'where the CMOS circuit has memory for storage Identification data for ECL probes in each of these hybrid chambers. φ GAS 136.9 preferably the volume of the hybridization chambers is less than 900,000 cubic microns. GAS136.10 Preferably, the hybridization chambers have a volume of less than 200,000 cubic microns. GAS136.il Preferably, the communication interface is a universal serial bus (USB) connector such that the housing is configured as a USB flash drive. GAS136.12 Preferably, the detection light sensor registers the detection photodiode array of the hybrid chambers. #GAS136.13 Preferably, the portable test module also has at least one calibration source for providing a calibration emission, and at least one calibration photodiode for sensing the calibration emission, wherein the CMOS circuit has A differential circuit subtracts the calibrated photodiode output from the output of one or more of the detection photodiodes. GAS136.14 Preferably, the portable test module also has a plurality of calibration sources and a plurality of corresponding calibration photodiodes, the calibration photodiodes being respectively registered with the calibration source. GAS136.15 Preferably, the calibration source is a calibration probe that does not have an ECL luminophore -183-201211533. GAS136.16 Preferably, the portable test module also has a plurality of calibration chambers, the calibration chamber containing calibration sources distributed throughout the array of hybridization chambers, wherein when used, from the detection photodiode The output of either one is compared to the output from the calibrated photodiode closest to the detection photodiode. GAS136.17 Preferably, the calibration source is a calibration probe and the calibration chamber is configured to seal the calibration probe from the fluid containing the target nucleic acid sequence. GAS 136.18 Preferably, each of the calibration chambers is surrounded by a three by three hybrid cell block. GAS136.19 Preferably, the distance between the detection photodiode and the hybridization chamber is less than 1,600 microns. GAS 1 3 6.20 Preferably, the ECL probe has a stem-loop structure, the loop portion of which contains a sequence complementary to the target nucleic acid sequence, the loop portion being located in the function for quenching photon emission from the ECL luminophore Between the sexual moiety and the ECL luminophore, such that hybridization with the target nucleic acid sequence opens the loop moiety and moves the ECL luminophore away from the functional moiety. The easy-to-use, mass-produced, inexpensive, lightweight, and portable test module accepts biological samples, using its integrated imaging sensor and integrated driver for exciting electrochemiluminescence to electrochemiluminate Probe hybridization identifies the nucleic acid sequence of the sample and provides an electronic result at the output of the probe. The electrochemiluminescence-based detection analysis eliminates the need for a detection system that requires a light source, excitation optics, and optical filter components to provide a lighter and less expensive inspection system. It is not necessary to reject any excitation light and simplify the detector circuit, making the detection system even cheaper. GAS 13 7.1 Aspects of the invention provide a test module for exciting an electrochemiluminescent probe configured to detect a target nucleic acid sequence, the test module comprising: a housing having a container for accepting the inclusion a fluid of the target nucleic acid sequence; an electrochemiluminescence (ECL) probe having an ECL luminophore that emits photons when in an excited state and a functional moiety that quenches photon emission from the ECL luminophore by resonance energy transfer; An electrode for receiving an electrical pulse to excite the ECL luminophore; a detection light sensor for exposing to photons emitted by the ECL luminophore; a control circuit for supplying electrical pulses to the electrodes; and a φ versatility string a busbar (USB) connector such that the housing is configured as a USB flash drive for transmitting information about the detection of the fluid to the external device; wherein, when in use, the nucleic acid sequence has been detected One of the ECL probes is reconfigured such that the functional portion does not quench photon emission from the ECL luminophore when excited by the electrodes. Preferably, the electrodes are sheets of electrically conductive material, the edge characteristics of the plates being configured such that the perimeter edges of each of the plates are greater than 128 microns. -185-201211533 GAS137·3 Preferably, the test module also has a lab-on-a-chip (LOC) device in which the ECL probe, electrode, detection light sensor and control circuit are integrated into the LOC device, wherein The LOC device has a support substrate for supporting the control circuit, which in turn supports the detection light sensor, electrodes, and ECL probes. GAS 137.4 Preferably, the control circuit is configured to provide electrical pulses to the layers of the CMOS circuits of the electrodes. GAS 137.5 Preferably, the electrical pulse has a period of less than 0.69 seconds. Preferably, the electrical current of the electrical pulse is between 0.1 nanoamperes and 69.0 nanoamperes. GAS 137.7 Preferably, the test module also has an array of hybridization chambers containing ECL probes for a different nucleic acid sequence and a pair of electrodes, wherein the control circuit has a memory for storage Identification data for ECL probes in each of these hybrid chambers. GAS 1 37.8 Preferably, the C Μ Ο S circuit is configured to apply a voltage to an electrode pair in each of the hybridization chambers. The voltage system is between 1.7 volts and 2.8 volts. GAS 137.9 Preferably, the voltage is between 1.9 volts and 2.6 volts. GAS 1 37.1 0 Preferably, the volume of the hybridization chambers is less than 900,000 cubic microns. GAS137.il Preferably, the hybridization chambers have a volume of less than 200,000 cubic microns. GAS137.12 Preferably, the detection light sensor is configured to register the detection photodiode array of the hetero-186-201211533 communication chamber. GAS 137.13 Preferably, the test module also has: at least one calibration source for providing a calibration emission and a calibration light sensor for sensing the calibration emission, wherein the control circuit has a differential circuit for Detecting the light sensor output minus the calibration light sensor output. Preferably, the test module also has a plurality of calibration sources, wherein the detection light sensor is respectively configured with a photodiode array of each of the ECL probes, and the calibration light sensor system A plurality of calibrated photodiodes respectively registered with the calibration sources. GAS 1 37.1 5 Preferably, the calibration source is a calibration probe that does not have an ECL luminophore. Preferably, the test module also has a plurality of calibration chambers containing calibration sources distributed throughout the array of hybridization chambers, wherein when used, from any of the detection photodiodes The output of the person is compared with the output of the calibrated photodiode from the closest photodetector. GAS 137.1. Preferably, the calibration source is a calibration probe and the calibration chamber is configured to block the calibration probe from the fluid containing the target nucleic acid sequence. GAS 137.18 Preferably, each of the calibration chambers is surrounded by a three by three hybrid cell block. GAS 1 37.19 Preferably, the distance between the detection photodiode and the hybridization chamber is less than 1,600 microns. GAS 1 3 7.20 Preferably 'the ECL probe has a stem-loop structure, -187-201211533, the loop portion of which contains a sequence complementary to the target nucleic acid sequence, the loop portion being located for quenching from the ECL luminophore Between the functional portion of the photon emission and the ECL luminophore, such that hybridization with the target nucleic acid sequence opens the loop portion and moves the ECL luminophore away from the functional portion. The easy-to-use, mass-produced, inexpensive, lightweight, and portable test module accepts biological samples, using its integrated imaging sensor and integrated driver for exciting electrochemiluminescence to electrochemiluminate The probe hybridizes to identify the nucleic acid sequence of the sample and provides an electronic result at the output of the probe, which is used to supply the module power and communication requirements. The electrochemiluminescence-based detection analysis eliminates the need for detection systems that require excitation light sources, excitation optics, and optical filter elements to provide a lighter and less expensive inspection system. It is not necessary to reject any excitation light and simplify the detector circuit, making the detection system even cheaper. GAS 1 3 8 · 1 This aspect of the invention provides a test module for detecting a target nucleic acid sequence in a fluid, the test module comprising: an outer casing having an inlet for receiving a nucleic acid sequence containing the target a fluidization chamber installed in the housing, the hybridization chamber comprising electrochemiluminescence (ECL) probes for detecting the target nucleic acid sequence, each of the ECL probes having a photon for emitting in an excited state An ECL luminophore and a functional moiety for quenching photon emission from the ECL luminophore by resonance energy transfer; an electrode for receiving an electrical pulse to excite the ECL luminophore; and -188-201211533 reagent reservoir, It contains an agent for addition to the fluid prior to detecting the target nucleic acid sequence; wherein the hybridization chamber has a volume of less than 900,000 cubic microns; and the reagent reservoir has a volume of less than 1,000,000,000 cubic microns. GAS 138.2 Preferably, the hybridization chamber has a volume of less than 200,000 cubic microns and the reagent reservoir has a volume of less than 300,000,000 cubic micrometers. φ GAS 138.3 Preferably, the hybridization chamber has a volume of less than 40,000 cubic microns and the reagent reservoir has a volume of less than 7 Å, 〇〇〇, 〇〇〇 cubic micrometers. GAS 138.4 Preferably, the hybridization chamber has a volume of less than 9,000 cubic microns and the reagent reservoir has a volume of less than 20,000,000 cubic microns. GAS 138.5 Preferably, the test module also has: a detection light sensor for exposing to photons emitted by the ECL luminophore; and a control circuit for supplying electrical pulses to the electrodes. GAS 13 8.6 Preferably, the test module also has: a communication interface of the control circuit for transmitting data to an external device. GAS 138.7 Preferably, the test module also has an array of hybridization chambers containing ECL probes and a pair of electrodes for different target nucleic acid sequences, wherein the control circuit has a body for storage Identification data for ECL probes in each of these hybrid chambers. GAS 1 3 8 · 8 Preferably, the communication interface is a universal serial confluence -189 - 201211533 row (USB) connector such that the housing is configured as a USB flash drive. GAS 138.9 Preferably, the detection light sensor registers the detection photodiode arrays of the hybrid chambers. GAS138.10 Preferably, the test module also has a lab-on-a-chip (LOC) device having the hybrid chamber, reagent reservoir, detection photodiode, electrode and ECL probe, the LOC device A support substrate having a layer of a CMOS circuit that forms part of the control circuit such that the detection photodiode is interposed between the hybridization chamber and the CMOS circuit.

GAS 138.il Preferably, the test module also has a plurality of reagent reservoirs and a polymerase chain reaction (PCR) portion that are inserted into the LOC device, wherein the PCR portion is configured to amplify the target nucleic acid sequence and The reagent reservoir contains a polymerase, dNTPs, and primers. GAS 138.12 Preferably, the LOC device has an upper cover to which the reagent reservoir is defined. GAS 1 3 8 . 1 3 Preferably, each of the reagent reservoirs has a surface tension valve, each of the surface tension valves having a meniscus anchor for forming a meniscus to retain reagent therein. GAS 138.14 Preferably, the electrode pairs in each of the hybridization chambers have an anode and a cathode respectively having a finger-like configuration, the finger structures being configured such that the fingers of the anode intersect the fingers of the cathode . GAS 138.15 Preferably, the CMOS circuit is configured to provide an electrical pulse to each of the pairs of electrodes to generate a voltage at the pair of electrodes, the voltage being between 1.7 volts and 2.8 volts. GAS 138.16 Preferably, the voltage is between 1.9 volts and 2.6-190-201211533 volts. GAS 138·17 Preferably, the distance between the detection photodiodes and the hybrid chambers is less than 1,600 microns and the calibration probes comprise functional portions for quenching photon emission. GAS138.18 Preferably, the ECL probe has a stem-loop structure, the loop portion of which contains a sequence complementary to the target nucleic acid sequence, the loop portion being located in the functionality for quenching photon emission from the ECL luminophore The portion φ is separated from the ECL luminophore such that hybridization with the target nucleic acid sequence opens the loop portion and moves the ECL luminophore away from the functional portion. GAS 138.19 Preferably, the electrical pulse transmitted to the electrode is a DC pulse and has a period of less than 0.69 seconds. GAS 1 3 8.20 Preferably, the electrical current of the electrical pulse is between 0.1 nanoamperes and 69.0 nanoamperes. The low volume hybridization chamber and reagent reservoir represent, to some extent, low probe and reagent volumes, thereby providing low probe and reagent cost and inexpensive detection of the φ system. The electrochemiluminescence-based detection analysis eliminates the need for detection systems that require excitation light sources, excitation optics, and optical filter elements to provide a lighter and less expensive inspection system. It is not necessary to reject any excitation light and simplify the detector circuit, making the detection system even cheaper. GAS 139.1 Aspects of the invention provide a test module for exciting an electrochemiluminescent probe configured to detect a target nucleic acid sequence, the test module comprising: an outer casing having an inlet for accepting the inclusion of the target Nucleic Acids - 191 - 201211533 Sequence Fluids; Electrochemiluminescence (ECL) Probes' have the functionality to emit photons of ECL luminescence when excited and the ability to quench photon emission from the ECL luminescence by resonance energy transfer a portion; and an electrode for receiving an electrical pulse to excite the ECL luminophores such that the ECL probe of the one of the labeled nucleic acid sequences has been reconfigured such that the functional portion is not quenched from The ECL luminophore emits photons when excited by the electrodes; wherein the ECL luminophore has a transition metal-coordination complex. GAS 139.2 Preferably, the transition metal-coordination complex is a ruthenium chelate. GAS 139.3 Preferably, the test module also has a detection light sensor for exposing photons emitted by the ECL luminophore; and a control circuit for supplying electrical pulses to the electrodes. GAS 139.4 Preferably, the test module also has a lab-on-a-chip (LOC) device, wherein the ECL probe, electrode, detection light sensor, and control circuitry are integrated into the LOC device, wherein the LOC device has A support substrate supporting the control circuit, which in turn supports the detection light sensor, the electrode, and the ECL probe. GAS 139.5 Preferably, the control circuit is configured to provide electrical pulses to the layers of the CMOS circuits of the electrodes. GAS 139.6 Preferably, the electrical pulses have a period of less than 0.69 seconds. GAS 139.7 Preferably, the electrical current of the electrical pulse is between 0.1 Na -192 - 201211533 amps to 69.0 Nai amps. GAS 13 9.8 Preferably, the test module also has: a communication interface of the control circuit for transmitting data to an external device. GAS 139.9 Preferably, the test module also has an array of hybridization chambers containing ECL probes for different target nucleic acid sequences, wherein the control circuit has memory for storage regarding each of the hybrids Identification data for indoor ECL probes. φ GAS 139.10 Preferably, the volume of the hybridization chamber is less than 900,000 cubic microns. GAS 139.il Preferably, the communication interface is a universal serial bus (USB) connector such that the housing is configured as a USB flash drive. GAS 139.12 Preferably, the detection light sensor registers the detection photodiode array of the hybrid chambers. GAS139.13 Preferably, the test module also has at least one calibration source for providing a calibration emission, and a calibration light sensor for sensing the φ calibration emission, wherein the control circuit has a differential circuit for use The calibration light sensor output is subtracted from the detected light sensor output. GAS139.14 Preferably, the test module also has a plurality of calibration sources, wherein the detection light sensor is respectively configured with a photodiode array registered with each of the ECL probes, and the calibration light sensor system is more A calibrated photodiode that is registered with the calibration sources, respectively. GAS 139.15 Preferably, the calibration source is a calibration probe that does not have an ECL luminophore. GAS 139.1 6 Preferably, the test module also has a plurality of calibration chambers - 193 - 201211533, which are distributed throughout the array of hybridization chambers, when used from the detection photodiode A calibration photoelectric photo from the closest to the detection photodiode. GAS 139.17 Preferably, the calibration source calibration chamber is configured to seal the calibration probe from containing the target fluid. GAS 139.18 Preferably, each of the calibration chambers is surrounded by a chamber block. GAS 13 9.19 Preferably, the distance between the detection photodiodes is less than 1,600 microns. GAS 1 3 9.2 0 Preferably, the ECL probe has a loop portion having a sequence complementary to the target nucleic acid sequence located between the photon emission from the ECL luminophore and the ECL luminophore Using the long-lived transition metal-coordination complex bottom probe with the loop portion of the target nucleic acid and the ECL luminophore away from the functional portion enhances the sensitivity and reliability of the detection system. The detection and analysis of the target of electrochemiluminescence is a lighter and cheaper detection system for detection of light source, excitation optics and optical filter elements. There is no need to reject the detector circuit, making the detection system even cheaper. GAS 140.1. The aspect of the invention provides a calibration source for the electrochemiluminescent probe for detecting the target nucleic acid sequence, the output of which is The hybrid with the output of the polar body and the three-by-three hybrid of the calibrated nucleic acid sequence and the hybridization chamber have a stem-loop structure, and the loop portion of the functional portion of the loop is hybridized to open the fraction. The electrochemiluminescence-based exemption requires any excitation, and thus provides excitation light and is also simple to excite the configured test module. The test-194-201211533 test module includes: an outer casing having an inlet for accepting the inclusion a fluid of a target nucleic acid sequence; an electrochemiluminescence (ECL) probe having an ECL luminophore that emits photons when in an excited state and a functional moiety that quenches photon emission from the ECL luminophore by resonance energy transfer; An electrode for receiving an electrical pulse to excite the ECL luminophores such that φ the ECL probe that has detected one of the labeled nucleic acid sequences is reconfigured such that the functional portion is not quenched from the ECL A photon is emitted when the luminophore is excited by the electrodes; wherein the ECL luminophore has a lanthanide metal-coordination complex. Preferably, the lanthanide metal-coordination complex is selected from the group consisting of: ruthenium chelate, ruthenium chelate or φ ruthenium chelate. GAS 140.3 Preferably, the test module also has a detection light sensor for exposing photons emitted by the ECL luminophore; and a control circuit for supplying electrical pulses to the electrodes. GAS 140.4 Preferably, the test module also has a lab-on-a-chip (LOC) device, wherein the ECL probe, electrode, detection light sensor and control circuit are integrated into the LOC device, wherein the LOC device has A support substrate supporting the control circuit further supports the detection light sensor, the electrode, and the ECL probe. - 195 - 201211533 GAS 140.5 Preferably, the control circuit is configured to provide electrical pulses to the layers of the CMOS circuits of the electrodes. GAS 140.6 Preferably, the electrical pulse has a period of less than 0.69 seconds. Preferably, the electrical current of the electrical pulse is between 0.1 nanoamperes and 69.0 nanoamperes. GAS 140.8 Preferably, the test module also has: a communication interface of the control circuit to transmit data to an external device. GAS 140.9 Preferably, the test module also has an array of hybridization chambers containing ECL probes for different target nucleic acid sequences, wherein the control circuit has memory for storage regarding each of the hybrids Identification data for indoor ECL probes. GAS 140.10 Preferably, the hybridization chambers have a volume of less than 900,000 cubic microns. GAS 140.il Preferably, the communication interface is a universal serial bus (USB) connector such that the housing is configured as a USB flash drive. GAS 140.12 Preferably, the detection light sensor registers the detection photodiode arrays of the hybrid chambers. GAS 140.13 Preferably, the test module also has: at least one calibration source for providing a calibration emission, and a calibration light sensor for sensing the calibration emission, wherein the control circuit has a differential circuit for Detecting the light sensor output minus the calibration light sensor output. GAS 140.14 Preferably, the test module also has a plurality of calibration sources, wherein the detection light sensor is respectively associated with each of the ECL probes, the light-196-201211533 electric diode array, and the calibration light The sensor is a plurality of calibrated photodiodes respectively registered with the calibration sources. GAS 140.15 Preferably, the calibration source is a calibration probe that does not have an ECL luminophore. GAS 140.16 Preferably, the test module also has a plurality of calibration chambers, the calibration chamber containing calibration sources distributed throughout the array of hybridization chambers, wherein, when used, from any of the detection photodiodes The output of the comparator is compared to the output of φ from the calibration photodiode closest to the detection photodiode. GAS 140.17 Preferably, the calibration source is a calibration probe and the calibration chamber is configured to seal the calibration probe from the fluid containing the target nucleic acid sequence. GAS 140.18 Preferably, each of the calibration chambers is surrounded by a three by three hybrid cell block. GAS 140.19 Preferably, the distance between the detection photodiode and the hybridization chamber φ is less than 1,600 microns. GAS 140.20 Preferably, the ECL probe has a stem-loop structure, the loop portion of which contains a sequence complementary to the target nucleic acid sequence, the loop portion being located in the functional portion for quenching photon emission from the ECL luminophore Between the ECL luminophore and the nucleic acid sequence of the target, the loop portion is opened and the ECL luminophore is moved away from the functional portion. The use of the long-lived lanthanide metal-coordination complex electrochemiluminescent substrate probe enhances the sensitivity and reliability of the detection system. The electrochemiluminescence-based detection analysis eliminates the need for a detection system that requires a light source, excitation optics, and optical filter components to provide a lighter and less expensive inspection system. It is not necessary to reject any excitation light and simplify the detector circuit, making the detection system even cheaper. GAS 141.1 This aspect of the invention provides a test module for detecting a target nucleic acid sequence in a fluid, the test module comprising: an outer casing having an inlet for receiving a fluid containing the target nucleic acid sequence An electrode pair for receiving an electrical pulse; an electrochemiluminescence (ECL) probe adjacent to each of the pair of electrodes, each of the ECL probes having an ECL luminophore for emitting photons in an excited state and for borrowing An ECL probe from a functional portion of photon emission of the ECL luminophore is quenched by resonance energy transfer such that an electrical pulse transmitted to the pair of electrodes excites the ECL luminophore; wherein the ECL probes are not immobilized The surface forms a suspension in the fluid containing the target nucleic acid sequence during the assay. Preferably, GAS 141.2 has a spotted volume of less than 200,000 cubic microns each. GAS 14 1.3 Preferably, the volume of the liquid is less than 30,000 cubic microns each. GAS 141.4 Preferably, the volume of the liquid is less than 2,000 cubic microns each. GAS 141.5 Preferably, the ECL luminophore has a transition metal-coordination complex. GAS 141.6 Preferably, the test module also has: -198-201211533 a detection light sensor for exposing photons emitted by the ECL luminophore; and a control circuit for supplying electrical pulses to the electrodes. GAS 141.7 Preferably, the test module also has: the communication interface of the control circuit to transmit data to the external device. GAS 1 4 1.8. Preferably, the test module also has an array of hybridization chambers containing ECL probes for different target nucleic acid sequences, φ wherein the control circuit has memory for storage Identification data for ECL probes in each of these hybrid chambers. Preferably, the volume of the hybridization chambers is less than 900,000 cubic microns. GAS 141.10 Preferably, the ECL luminophore has a lanthanide metal-coordination complex. GAS 141.il Preferably, the communication interface is a universal serial bus (USB) connector such that the housing is configured as a USB flash drive. φ GAS141.12 Preferably, the detection light sensor registers the detection photodiode array of the hybrid chambers. GAS 14 1.1 3 Preferably, the test module also has: at least one calibration source for providing a calibration emission and a calibration light sensor for sensing the calibration emission, wherein the control circuit has a differential circuit for The calibration light sensor output is subtracted from the detected light sensor output. GAS 1 4 1 . 1 4 Preferably, the test module also has a plurality of calibration sources, wherein the detection light sensor is respectively configured with a photodiode array registered with each of the ECL probes, and the calibration light is The sensor is a plurality of calibrated photodiodes respectively registered with the calibration -199 - 201211533 source. GAS 14 1.1 5 Preferably, the calibration source does not have a calibration probe. GAS 1 4 1 . 16 Preferably, the test module also has a calibration chamber containing a plurality of detectors distributed throughout the array of hybridization chambers, and any one of the photodiodes from the detection is from the most The calibration photodiode is closer to the detection photodiode. GAS 14 1.1 7 Preferably, the calibration source is a calibration laboratory configured to seal the calibration probe from the fluid containing the target. GAS 141.1 8 Preferably, each of the calibration chambers is surrounded by three chambers. GAS 14 1.1 9 Preferably, the distance between the detection photodiodes is less than 1,6.00 microns. Preferably, the ECL probe has a loop portion having a sequence complementary to the target nucleic acid sequence, and is located between the photon emission component from the ECL luminophore and the ECL luminophore for quenching The target nucleic acid sequences the loop portion and leaves the ECL luminophore away from the functional portion. The suspended probes increase the sensitivity of reading by their deep electrical excitation depth. The suspended probe can be relatively sampled. The electrochemiluminescence-based detection analysis analyzes multiple calibration chambers of the ECL luminescence group, wherein the output system is the output ratio of the body and the calibration nucleic acid sequence By the hybridization of the three hybrids and the stem-loop structure of the hybrid chamber, the functional part of the loop is hybridized to open the 〇 and the large-emitting light is easy and inexpensive to remove. -200-201211533 light source, excitation optical and optical filter elements The detection system provides a lighter and less expensive inspection system. It is not necessary to reject any excitation light and simplify the detector circuit, making the detection system even cheaper. GAS 142.1 This aspect of the invention provides a test module for detecting a target nucleic acid sequence in a fluid, the test module comprising: an outer casing having an inlet for receiving a fluid containing the target nucleic acid sequence; An array of φ electrochemiluminescence (ECL) probes for hybridization with the target nucleic acid sequence to form a probe-target hybrid; and an electrode placed for receiving electrical pulses, the probe-target hybridization The system is configured to emit photons of light when excited by current between the electrodes: and a light sensor for detecting light emitted by the probes; wherein when used, the fluid is added to the The probe prevents subsequent addition of other fluids to the φ probes. GAS 142.2 Preferably, the test module also has an array of hybridization chambers, wherein each of the chambers comprises a pair of electrode pairs and a probe for one of the targets, wherein the fluid is acted upon by capillary action Fill each of these rooms. Preferably, the GAS 142.3 has a volume of less than 900,000 cubic microns each. GAS 142.4 Preferably, the test module also has a lab-on-a-chip (LOC) device, wherein the ECL probe, the electrode and the detection light sensor are integrated into the LOC device, wherein the LOC device has a CMOS circuit, 201 - 201211533 A support substrate for supporting the CMOS circuit, which in turn supports the detection light sensor, the electrode, and the ECL probe. ' GAS 142.5 Preferably, each of the ECL probes has an ECL luminophore for emitting photons in an excited state and a functional portion for quenching photon emission from the ECL luminophore by resonance energy transfer, The electrical pulse transmitted to the pair of electrodes is excited to excite the ECL luminophore. GAS 142.6 Preferably, the ECL probe has a stem-loop structure, the loop portion of which contains a sequence complementary to the target nucleic acid sequence, the loop portion being located in the functionality for quenching photon emission from the ECL luminophore Portion between the portion and the ECL luminophore to hybridize with the target nucleic acid sequence to open the loop portion and to move the ECL luminophore away from the functional portion. GAS 142.7 Preferably, the hybridization chambers each have a wall portion that is transparent to the light emitted by the probe. GAS 142.8 Preferably, the CMOS circuit is provided with the light sensor, wherein the wall portion is located between the probes and the light sensor. GAS 142.9 Preferably, the wall is doped with a layer of cerium oxide. GAS 142.10 Preferably, the CMOS circuit is configured to provide electrical pulses to the electrodes. GAS 142.il Preferably, the CMOS circuit is configured to apply a voltage to an electrode in each of the hybridization chambers, the voltage being between 1.7 volts and 2.8 volts. GAS 142.12 Preferably, the voltage is between 1.9 volts and 2.6 volts. GAS 142.13 Preferably, the electrical pulse has a 201211533 period of less than 0.69 seconds. GAS 142.14 Preferably, the electrical current of the electrical pulse is between 0.1 nanoamperes and 69.0 nanoamperes. GAS 142.15 Preferably, the electrodes have an anode and a cathode, each having a finger configuration, the finger structures being configured to intersect the fingers of the anode with the fingers of the cathode. Preferably, the anode and the cathode are separated by a dielectric gap of 0.4 φ micrometers to 2 micrometers wide. GAS 142.17 Preferably, the LOC device has a flow path configured to attract fluid containing the target to all of the hybrid chambers by capillary action. GAS 142.18 Preferably, the light sensor is an array of photodiodes locating the hybridization chamber such that each of the hybridization chambers corresponds to one of the photodiodes. Preferably, the ECL luminophores each have a lanthanide metal genus-coordination complex. GAS 1 42.20 Preferably, the ECL luminophores each have a transition metal-coordination complex. The easy-to-use, mass-produced, inexpensive and lightweight genetic test module accepts a biological sample, uses its integrated imaging sensor to identify the nucleic acid sequence of the sample via probe hybridization and provides electronic results at the output of the sample.检测The detection and analysis of the target based on electrochemiluminescence eliminates the need for a detection system that requires excitation of light sources, excitation optics and optical filter elements, thereby providing a lighter and cheaper detection system of -203- 201211533. It is not necessary to reject any excitation light and simplify the detector circuit, making the detection system even cheaper. GAS 1 43.1 This aspect of the invention provides a test module for amplifying and detecting a target nucleic acid sequence in a fluid, the test module comprising: a housing having a container for receiving a nucleic acid containing the target a fluid of sequence; a polymerase chain reaction (PCR) portion for amplifying the nucleic acid sequence of the target; an array of electrochemiluminescence (ECL) probes for hybridizing to the target nucleic acid sequence to form a probe a target hybrid; and an electrode placed for receiving an electrical pulse, the probe-target hybridization system being configured to emit photons of light when excited by current between the electrodes; and for detecting Light sensors of the light emitted by the probes: wherein, in use, the fluid is added to the probes to prevent subsequent addition of other fluids to the probes. GAS 143.2 Preferably, the test module also has an array of hybridization chambers, wherein each of the chambers comprises a pair of electrode pairs and a probe for one of the targets, wherein the fluid is acted upon by capillary action Fill each of these rooms. GAS 143.3 Preferably, the test module also has a plurality of reagent reservoirs in fluid communication with the PCR portion, wherein the reagent reservoirs comprise a polymerase, dNTPs, and primers. GAS 143.4 Preferably, the test module also has a lab-on-a-chip (LOC) device having a support substrate and a CMOS circuit, -204-201211533, wherein the LOC device has the ECL probe and the electrode The support substrate is supported by the CMOS circuit and the CMOS circuit has the detection light sensor adjacent to the electrode and the ECL probe. GAS 143.5 Preferably, each of the ECL probes has an ECL luminophore for emitting photons in an excited state and a functional portion for quenching photon emission from the ECL luminophore by resonance energy transfer, The electrical pulse transmitted to the pair of electrodes excites the ECL luminophore. φ GAS 143.6 Preferably, the ECL probe has a stem-loop structure, the loop portion of which contains a sequence complementary to the target nucleic acid sequence, the loop portion being located in the function for quenching photon emission from the ECL luminophore Between the sexual moiety and the ECL luminophore, such that hybridization with the target nucleic acid sequence opens the loop moiety and moves the ECL luminophore away from the functional moiety. GAS 143.7 Preferably, the hybridization chambers each have a wall portion that is transparent to the light emitted by the probe. GAS 143.8 Preferably, the wall is located between the probe and the photoreceptor. Preferably, the wall portion is entangled with a layer of cerium oxide. GAS 143.10 Preferably, the CMOS circuit is configured to provide electrical pulses to the electrodes. GASM3.il Preferably, the electrical pulses have a voltage between 1.7 volts and 2.8 volts. GAS143.12 Preferably, the voltage is between 1.9 volts and 2.6 volts. GAS 143.13 Preferably, the electrical pulse has a period of -205 - 201211533 of less than 0.69 seconds. GAS 143.14 Preferably, the current of the electrical pulse is between 0.1 Nai and 69.0 Nai's. GAS 143.15 Preferably, the electrodes have an anode and a cathode respectively having a finger-like structure, and the finger-like structures are It is configured such that the finger of the anode intersects the finger of the cathode. GAS 143.16 Preferably, the anode and cathode are separated by a dielectric gap of 0.4 microns to 2 microns wide. GAS 143.17 Preferably, the LOC device has a flow path configured to attract fluid containing the target to all of the hybrid chambers by capillary action. GAS 143.18 Preferably, the light sensor is an array of photodiodes locating the hybridization chamber such that each of the hybridization chambers corresponds to one of the photodiodes. GAS143.19 Preferably, the ECL luminophores each have a lanthanide metal-coordination complex. GAS 1 43.20 Preferably, the ECL luminophores each have a transition metal-coordination complex. The easy-to-use, mass-produced, inexpensive and lightweight genetic test module accepts a biological sample, amplifies the nucleic acid target in the sample, and uses the integrated imaging sensor to identify the nucleic acid sequence of the sample via probe hybridization. And provide electronic results at the output of the other. The electrochemiluminescence-based detection analysis eliminates the need for detection systems that require excitation light sources, excitation optics, and optical filter elements to provide a lighter and less expensive inspection system. It is not necessary to reject any excitation light and simplify the detector circuit, making the detection system even cheaper. GAS 144.1 This aspect of the invention provides a test module for detecting a target nucleic acid sequence in a fluid, the test module comprising: a housing having a container for receiving a fluid containing the target nucleic acid sequence; An array of electrochemiluminescence (ECL) probes for hybridizing to the target nuclear φ acid sequence to form a probe-target hybrid; and an electrode placed for receiving electrical pulses, the probes - The target hybridization system is configured to emit photons when excited by current between the electrodes; and a control circuit for providing electrical pulses to the electrodes; wherein, when in use, the fluid is added to the probes to prevent subsequent Add other fluids to the probes. φ GAS 144.2 Preferably, the test module also has: a light sensor for detecting photons emitted by the probes; and an array of hybridization chambers, wherein each of the chambers comprises a pair of electrodes and a probe of one of the targets; wherein the fluid is charged to each of the chambers by capillary action. Preferably, the GAS 144.3 has a volume of less than 900,000 cubic microns each. GAS 144.4 Preferably, the test module also has a lab-on-a-chip (LOC) device having a support substrate, a CMOS circuit, the 207-201211533, the EC L probe, the electrode, and the light sensor, wherein The support substrate supports a CMOS circuit having the detection light sensor, which in turn supports the electrode and the ECL probe. GAS 144.5 Preferably, each of the ECL probes has an ECL luminophore for emitting photons in an excited state and a functional portion for quenching photon emission from the ECL luminophore by resonance energy transfer, The electrical pulse transmitted to the pair of electrodes excites the ECL luminophore. GAS 144.6 Preferably, the ECL probe has a stem-loop structure, the loop portion of which contains a sequence complementary to the target nucleic acid sequence, the loop portion being located in the functionality for quenching photon emission from the ECL luminophore Portion between the portion and the ECL luminophore to hybridize with the target nucleic acid sequence to open the loop portion and to move the ECL luminophore away from the functional portion. GAS 144.7 Preferably, the hybridization chambers each have a wall portion that is permeable to photon light emitted by the probe. GAS 144.8 Preferably, the CMOS circuit is provided with the light sensor, the wall being located between the probes and the light sensor. GAS 144.9 Preferably, the wall is entangled with a layer of cerium oxide. GAS 144.10 Preferably, the CMOS circuit is configured to provide electrical pulses to the electrodes. GAS 144.il Preferably, the electrical pulse has a voltage between 1.7 volts and 2.8 volts. GAS 144.12 Preferably, the voltage is between 1.9 volts and 2.6 volts. GAS 1 44.1 3 Preferably, the electrical pulse has a period of -208-201211533 less than 〇.69 seconds. GAS 144.14 Preferably, the electrical current of the electrical pulse is between 0.1 nanoamperes and 69.0 nanoamperes. GAS 144.15 Preferably, the electrodes have an anode and a cathode, each having a finger configuration, the finger structures being configured to intersect the fingers of the anode with the fingers of the cathode. GAS 144.16 Preferably, the anode and cathode are separated by a dielectric gap of 0.4 φ micrometers to 2 micrometers wide. GAS 144.17 Preferably, the LOC device has a flow path configured to attract fluid containing the target to all of the hybridization chambers by capillary action. GAS 144.1 8 Preferably, the light sensor is an array of photodiodes locating the hybridization chamber such that each of the hybridization chambers corresponds to one of the photodiodes. GAS 144.19 Preferably, the ECL luminophores each have a lanthanide genus-coordination complex. GAS 144.20 Preferably, the ECL luminophores each have a transition metal-coordination complex. The easy-to-use, mass-produced, inexpensive and lightweight genetic test module accepts a biological sample and uses an integrated imaging sensor with a driver for exciting the electroluminescent luminophore to identify the sample via probe hybridization. The nucleic acid sequence, and at its output, provides an electronic result. The electrochemiluminescence-based detection analysis eliminates the need for detection systems that require excitation light sources, excitation optics, and optical filter elements to provide a lighter and less expensive inspection system. It is not necessary to reject any excitation light and simplify the detector circuit, making the detection system even cheaper. GAS 146.1 This aspect of the invention provides a wafer-on-lab (LOC) device for detecting a target nucleic acid sequence in a fluid, the LOC device comprising: an electrochemiluminescence (ECL) probe for detecting the target nucleic acid sequence a needle, each of the probes having an ECL luminophore for emitting photons in an excited state, a functional portion for quenching photon emission from the ECL luminophore by resonance energy transfer; and for receiving an electrical pulse Electrodes that excite the ECL luminophores; wherein the electrodes are arranged in pairs, each pair having an anode and a cathode separated by a dielectric gap between 0.4 microns and 2 microns wide. GAS 146.2 Preferably, the LOC device also has a light sensor located adjacent to the probe for sensing those probes that respond to electrical pulses to produce photons. GAS 146.3 Preferably, the LOC device also has an array of hybridization chambers, wherein each of the chambers comprises a pair of electrode pairs and a probe for one of the targets, wherein the fluid is acted upon by capillary action Preferably, the LOC device also has a support substrate, wherein the light sensor is an array of photodiodes on the support substrate that are registered with the hybrid chambers. GAS 146.5 Preferably, the photodiode array is less than 1,600 microns from the probe. -210-201211533 GAS146.6 Preferably, the LOC device also has a CMOS circuit on the support substrate, the array of the photodiode being a component of the CMOS circuit, wherein the CMOS circuit responds when not in use An ECL photon from the positive control probe is measured to cause an error signal. GAS 146.7 Preferably, the LOC device also has at least one negative control chamber containing a negative control probe that is incapable of hybridizing to any nucleic acid sequence in the fluid. φ GAS146.8 Preferably, the LOC device also has: a flow path containing the target fluid; wherein the CMOS circuit is interposed between the hybridization chamber and the support substrate, and the flow path is acted upon by capillary action The fluid is attracted to each of the hybrid chambers. GAS 146.9 Preferably, the LOC device also has a lysis unit, wherein the flow system contains a biological sample of cells and the lysate destroys the cell membrane to release any genetic material therein. GAS 146.10 Preferably, the hybridization chambers each have a wall portion that is permeable to photon light emitted by the probe φ. Preferably, the wall portion is between the probe and the array of photodiodes. Preferably, the wall portion is doped with a layer of cerium oxide. GAS146.13 Preferably, the probe has a stem-loop structure, the loop portion of which contains a sequence complementary to the target nucleic acid sequence, the loop portion being located in the functional portion for quenching photon emission from the ECL luminophore Between the ECL luminophore and the nucleic acid sequence of the target, the loop portion is opened and the ECL luminophore is moved away from the functional portion. Preferably, the CMOS circuit is configured to provide electrical pulses to the electrodes. GAS 1 46.1 5 Preferably, the electrical pulse has a period of less than 〇 · 6.9 seconds. GAS 1 46.1 6 Preferably, the electrical current of the electrical pulse is between 0.1 nanoamperes and 69.0 nanoamperes. GAS 146.17 Preferably, the electrodes have an anode and a cathode, each having a finger configuration, the finger structures being configured to intersect the fingers of the anode with the fingers of the cathode. GAS 146.18 Preferably, the ECL luminophores each have a transition metal-coordination complex. GAS 146.19 Preferably, the LOC device also has a PCR portion for amplifying the target nucleic acid sequence prior to detection by the probes. GAS 1 46.20 Preferably, the LOC device also has an upper cover having a reagent reservoir for addition to the fluid prior to detecting the target sequence. The electrochemiluminescence-based detection analysis eliminates the need for detection systems that require excitation light sources, excitation optics, and optical filter elements to provide a lighter and less expensive inspection system. It is not necessary to reject any excitation light and simplify the detector circuit, making the detection system even cheaper. The need to add an electrochemical co-reactant makes the detection system simpler and less expensive, helping to use more of the detection chemistry options. GPC03 5.1 This aspect of the invention provides a wafer-on-lab (LOC) device for amplifying a target nucleic acid sequence in a sample, the LOC-212-201211533 comprising: a support substrate; a plurality of processing the sample The function portion 'of the functional portions is a nucleic acid amplification portion for amplifying a nucleic acid sequence in a sample; wherein the nucleic acid amplification portion is supported by the support substrate' and the support substrate is defined for thermal isolation A groove between one or more of the nucleic acid amplification unit and another functional unit. φ GPC035.2 Preferably, one of the functional portions is provided with a hybridization portion of the probe array for hybridizing with the target nucleic acid sequence and the groove is extended in the nucleic acid amplification portion and the hybridization portion The heat generated by the hybridization portion and the nucleic acid amplification portion is thermally isolated. GPC03 5.3 Preferably, the support substrate is a layer of a substance having a layer separated by a thickness of the substrate, the functional portions being formed on one surface of the support substrate and the groove being attached to the other Surface definition. GPC035.4 Preferably, the LOC device also has a CMOS circuit supported by the support φ substrate, wherein the trench extends through the substrate to the CMOS circuit GPC03 5.5. Preferably, the trench contains air during use. GPC035.6 Preferably, the nucleic acid amplification unit is a polymerase chain reaction (PCR) unit. GPC035.7 Preferably, the LOC device also has temperature sensing

And a microsystem technology (MST) layer having the functional portion, wherein the CMOS circuit is between the support substrate and the M ST layer, the CMOS circuit configured to use the temperature sensor output to provide feedback Control the PCR -213 - 201211533. GPC03 5.8 Preferably, the PCR portion has a PCR microchannel for heating to circulate the sample, the PCR microchannel defining a flow path having a cross-sectional area of less than 1 000,000 square microns. GPC03 5.9 Preferably, the PCR microchannel has a cross-sectional area of less than 1 6,000 square microns perpendicular to the flow. GPC 03 5.1 0 Preferably, the PCR microchannel has a cross-sectional area of less than 2,500 square microns perpendicular to the flow. GPC03 5.il Preferably, the PCR microchannel has at least one elongated heater element extending parallel to the PCR microchannel. GPC03 5.1 2 Preferably, the PCR portion has a plurality of elongated PCR chambers respectively formed by respective portions of the PCR microchannels, the PCR microchannels having a curved configuration formed by a series of wide curves, each of which A wide curve forms the channel portion of one of the elongate PCR chambers. GPC03 5.1 3 Preferably, the PCR portion has an active valve for retaining liquid in the PCR portion during thermal cycling and allowing the liquid to flow to the hybridization chamber array in response to an activation signal from the CMOS circuit. GPC03 5.1 4 Preferably, the active valve is provided with a meniscus anchor and a boiling start valve of the heater, the meniscus anchor configured to form a meniscus to prevent capillary flow of the liquid The heater is used to boil the liquid to release the meniscus from the meniscus anchor to restore the capillary drive flow. GPC 0 3 5.1 5 Preferably, the LOC device also has a light sensor for detecting hybridization of any of the probes within the hybrid. -214-201211533 GPC03 5.1 6 Preferably, the nucleic acid amplification unit is a thermostatic nucleic acid amplification unit. GPC03 5.1 7 Preferably, the hybridization portion has a hybridization chamber array' that is adapted to contain probes such that the probes within each of the hybridization chambers are configured to hybridize to one of the target nucleic acid sequences. GPC03 5.1 8 Preferably, the light sensor is registered with the photodiode array of the hybridization chamber. GPC03 5.1 9 Preferably, the thermolabic nucleic acid amplification section has a nucleic acid amplification microchannel for maintaining a reaction temperature of the sample, the nucleic acid amplification microchannel defining a flow path having a cross-sectional area of the flow of less than 100,000 square micrometers. GPC03 5.20 Preferably, the nucleic acid amplification microchannel has a cross-sectional area of the stream of less than 1 6,000 square microns. The easy-to-use, mass-produced and inexpensive genetic analysis LOC device accepts a sample containing nucleic acid and then amplifies the nucleic acid target in the sample in the PCR chamber of the device. The PCR chamber is thermally insulated using an insulated trench, which provides rapid temperature cycling of the mixture, reduces power requirements, and reduces the effects of the heating cycle on other parts of the device, allowing for a broader range of detection chemistry. This rapid temperature cycling capability increases the speed of this detection. GPC03 6.1 This aspect of the invention provides a microfluidic device comprising: a sample inlet for receiving a sample of biological material having a nucleic acid sequence; and a nucleic acid amplification portion for amplifying the nucleic acid sequence, the nucleic acid amplification Additions - 215 - 201211533 have an amplification chamber and a heater element; wherein the heater element is bonded to the interior surface of the amplification chamber. GPC036.2 Preferably, the nucleic acid amplification portion is a polymerase chain reaction (PCR) portion and the amplification chamber is a PCR chamber such that the heater element is heated together to circulate the sample with dNTPs, primers, polymerases, and buffers. A solution to amplify the nucleic acid sequence. GPC036.3 Preferably, the microfluidic device also has a support substrate for supporting the PCR portion, wherein the PCR portion has a microchannel having a PCR inlet and a PCR outlet, the PCR chamber being part of the microchannel And the inner surface of the combined heater element is closest to the bottom of the microchannel of the support substrate. GPC036.4 Preferably, the PCR portion has a plurality of PCR chambers, and the microchannels have a curved configuration formed by a series of wide curves, each of the wide curve channel portions forming one of the PCR chambers By. GPC03 6.5 Preferably, each of the channel portions has a plurality of the heater elements. GPC03 6.6 Preferably, the plurality of heater elements are elongate in position aligned with the longitudinal axis of the channel portion and along the end to the end of the channel portion. GPC036.7 Preferably, each of the plurality of elongate heater elements is independently operable. GPC036.8 Preferably, the microfluidic device also has at least one temperature sensor for feedback control of the elongate heater element. GPC036.9 Preferably, the PCR portion has an active valve at the PCR outlet for retaining the liquid during the heating cycle within the PCR section of the -216-201211533. GPC036.1 0 Preferably, the active valve is provided with a boiling start valve of a meniscus anchor, the anchor is for retaining liquid in the PCR portion, and the boiling start valve also has a valve heater It is used to boil the liquid to release the meniscus from the meniscus anchor to recover the capillary drive and exit the PCR section. GPC03 6.il Preferably, the meniscus anchor is bored and the valve plus φ heater is located adjacent the edge of the bore. GPC036.1 2 Preferably, the PCR microchannel has a cross-sectional area of the fluid ranging from 400 square micrometers to 1 square micrometer. GPC03 6.1 3 Preferably, the microfluidic device also has a dialysis unit, wherein the biological material comprises cells of different sizes, the dialysis portion being configured to separate a sample larger than a predetermined valve cell to a portion of the sample system The treatment is performed separately from the remaining sample containing only cells smaller than the predetermined valve. G# GPC03 6.1 4 Preferably, the nucleic acid sequence is derived from cells smaller than the predetermined valve. GPC 036.1 5 Preferably, each of the channel portions has a liquid sensor proximate one end, the liquid sensor being configured to detect liquid at the liquid detector position to feedback control the heater element. GPC036.1 6 Preferably, the microfluidic device also has a reagent reservoir for containing reagents for nucleic acid amplification use, and a surface tension valve provided with a hole configured to cause the reagent to form a meniscus The surface is such that the meniscus retains the reagent in the reagent reservoir until the fluid sample contacts the surface of the meniscus and the reagent exits the reagent reservoir. GPC036.1 7 Preferably, the reservoir has a venting opening as an air inlet when the reagent exits the reagent reservoir. GPC036.1 8 Preferably, the microfluidic device also has a probe array for hybridizing with the target nucleic acid sequence in the sample to form a probe, and a hybrid for detecting the probe-target Light sensor. GPC036.1 9 Preferably, the PCR section has a thermal cycle time of less than 4 seconds. GPC036.20 Preferably, the PCR portion has a thermal cycle time of between 0.45 seconds and 1.5 seconds. The easy-to-use, mass-produced and inexpensive microfluidic device accepts a sample containing nucleic acid, and then amplifies the nucleic acid target in the sample using the nucleic acid amplification chamber of the device. GLY001.1 This aspect of the invention provides a microfluidic device for dissolving cells in a fluid, the microfluidic device comprising: an inlet for receiving a fluid containing cells; a lysis reagent reservoir containing a lysis reagent And an outlet valve: and a lysate in fluid communication with the inlet for retaining the fluid during cell lysis; wherein the outlet valve is located upstream of the lysis portion and configured to flow into the solution at the fluid The cell is opened, thereby adding the lysis reagent to the fluid. GLY001.2 Preferably, the outlet valve is provided with a surface tension valve of the bore configured to form a meniscus to retain the lysis reagent therein until the fluid is removed upon contact with the fluid The lysis reagent is added to the -218-201211533 fluid stream to enter the lysis section. GLY001.3 Preferably, the lysis portion is configured to attract a lytic channel of fluid from the inlet by capillary action, the lysis portion having an active valve at a downstream end of the lysis channel for use in The fluid is retained during cell lysis. GLY001.4 Preferably, the active valve is provided with a boiling start valve of a meniscus anchor, the anchor is used for retaining the fluid φ in the lysis unit, and the boiling start valve also has a valve heater It is used to boil the liquid to release the meniscus from the meniscus anchor to recover the capillary drive and flow out of the lysis unit. GLY001.5 Preferably, the cross-sectional area of the microchannel perpendicular to the fluid is between 8 square microns and 2,00 0 square microns. GLY001.6 Preferably, the microfluidic device also has a support substrate, wherein the inlet, the lysis unit, and the lysis reagent reservoir are supported by the support substrate and configured as a on-wafer laboratory (LOC) device. φ GLY001.7 Preferably, the microfluidic device also has a CMOS circuit between the support substrate and the lysis unit, the CMOS circuit being configured for operational control of the valve heater. GLY001.8 Preferably, the microfluidic device also has a polymerase chain reaction (PCR) portion provided with at least one heater for heating the circulating fluid to amplify the nucleic acid sequence released from the cells. GLY001.9 Preferably, the microfluidic device also has at least one sensor, wherein the CMOS circuit is configured to use the sensor to feedback control the at least one PCR heater. -219 - 201211533 GLY001.10 Preferably, the microfluidic device also has a plurality of heaters and a plurality of temperature sensors and a plurality of liquid sensors to cause the CMOS circuit to respond to the liquid sensor to control the The initial start of the heater and the response to the temperature sensor to control the heater power. GLY001.il Preferably, each of the plurality of heaters is independently operable. GLY00 1 . 1 2 Preferably, the PCR portion is configured as a PCR microchannel having a cross-sectional area of from 1 square micron to 400 square micrometers. The PCR portion also has an active valve located at a downstream end of the PCR microchannel. Used to retain the fluid during amplification of the nucleic acid sequence. GLY001.13 Preferably, the microfluidic device also has a PCR mixture reservoir and a polymerase reservoir, wherein the PCR mixture reservoir comprises dNTPs, primers, and a buffer solution and the polymerase reservoir comprises a polymerase. GLY001.14 Preferably, the microfluidic device also has an upper cover defining the lysis reagent reservoir, the PCR mixture reservoir, the polymerase reservoir and the lysis unit, and the PCR portion is interposed Between the upper cover and the support substrate. GLY001.15 Preferably, the microfluidic device also has a dialysis unit, wherein the cells in the fluid are different in size, and the dialysis portion is configured to separate a sample larger than a predetermined valve to a portion of the sample system. It is treated separately from the remaining sample containing only cells smaller than the predetermined valve. GLY001.16 Preferably, the nucleic acid sequence is derived from cells larger than the predetermined valve. GLY001.17 Preferably, the microfluidic device also has an anticoagulant-220-201211533 reservoir, wherein the flow system is a whole blood sample such that an anticoagulant is added to the whole blood upstream of the dialysis portion and the The dialysis section is configured to concentrate white blood cells to a portion of the whole blood sample. GLY001.18 Preferably, the microfluidic device also has a hybridization portion downstream of the PCR portion, the hybridization portion having a probe array for hybridization with a predetermined target nucleic acid sequence in an amplicon from the PCR portion; A light sensor for detecting probe hybridization in a probe array. φ GLY001.19 Preferably, the hybridization portion has an electrode placed to receive an electrical pulse, and the probe is an electrochemiluminescence (ECL) probe for hybridization with the target nucleic acid sequence to form a probe-target hybrid. A needle, the probe-target hybridization system is configured to emit photons of light when excited by current between the electrodes. GLY001.20 Preferably, the light sensor is positioned to respectively register the photodiode array of the ECL probe. The easy-to-use, mass-produced, inexpensive microfluidic device accepts a biochemical sample, uses a chemical lysis subunit to dissolve the cell or cell organelle, and treats the lysate. The lysis process is performed by cell extraction analysis and diagnostics in the sample, and subsequent processing and analysis of the targets are provided. Integrating the lysis subunit into the device provides a simple detection step, a low number of system components, and a simple system manufacturing process, resulting in an inexpensive detection system. GLY002.1 This aspect of the invention provides a microfluidic device for dissolving cells in a fluid, the microfluidic device comprising: an inlet channel for receiving a fluid containing cells; -221 - 201211533 being in fluid communication with the inlet channel Retaining the fluid between the lysis chambers; and heating the fluid in the lysis chamber with a heater; wherein the inlet passage is configured for passage. GLY002.2 Preferably, the lysing action retains the lysate of the fluid from the inlet to retain the fluid during active valve resection at the downstream end of the lysing microchannel. GLY002.3 Preferably, the active valve is a boiling start valve, the anchor is used for retention, and the boiling start valve also has a valve heater, the meniscus is released from the meniscus anchor, The lysis unit. GLY002.4 Preferably, the microchannel is between 1 square micron and 40 square micron. GLY002.5 Preferably, the microfluid, wherein the inlet and the lysis unit are supported by the on-wafer laboratory (LOC) device. GLY002.6 Preferably, the CMOS circuit between the microfluidic substrate and the lysis chamber is configured to operate a valve heater that controls the lysis heater and the point. Capsule capillary lysis for lysing the cells during cell lysis, the lysate is configured to be provided by a capillary channel, the lysate having a meniscus anchor for use in the cell solutes The flow system in the lysing portion is used to boil the liquid to cause the capillary to be driven to flow out of the cross-sectional area of the fluid. The device also has support substrate support and is configured to have the device also located in the branch. Located at the downstream of the microchannel -222-201211533 GLY002.7 Preferably, the microfluidic device also has a polymerase chain reaction (PCR) portion provided with at least one heater for heating and circulating the fluid for amplification Nucleic acid sequences released from such cells. GLY002.8 Preferably, the microfluidic device also has at least one sensor, wherein the CMOS circuit is configured to use the sensor to feedback control the at least one P C R heater. GLY002.9 Preferably, the microfluidic device also has a plurality of temperature φ sensors and a plurality of liquid sensors, wherein the PCR portion has a plurality of heaters to cause the CMOS circuits to respond to the liquid sensors for control The heater is initially activated and returned to the temperature sensor to control the heater power. GLY002.1 0 Preferably, each of the plurality of elongated heaters is independently operable. Preferably, the PCR portion is configured to have a PCR microchannel having the same cross section as the lysing microchannel, and the PCR portion also has an active valve located at a downstream end of the PCR microchannel to amplify the nucleic acid sequence #期 is used to retain the fluid. GLY002.12 Preferably, the microfluidic device also has a PCR mixture reservoir and a polymerase reservoir, wherein the PCR mixture reservoir comprises dNTPs, primers, and a buffer solution and the polymerase reservoir comprises a polymerase. GLY002.1 3 Preferably, the microfluidic device also has an upper cover, the upper cover defines the PCR mixture reservoir and the polymerase reservoir, wherein the lysis unit and the PCR portion are interposed between the upper cover and the support substrate between. GLY002.14 Preferably, the microfluidic device also has a dialysis unit, wherein the cells in the fluid are different in size, and the dialysis portion is configured to divide the cells larger than the predetermined valve to a part of the sample from -223 to 201211533, The portion of the sample is treated separately from the remaining sample containing only cells smaller than the predetermined valve. GLY002.1 5 Preferably, the nucleic acid sequence is derived from cells larger than the predetermined valve. GLY002.1 6 Preferably, the microfluidic device also has an anticoagulant reservoir, wherein the flow system is a whole blood sample such that an anticoagulant is added to the whole blood upstream of the dialysis portion and the dialysis system is It is configured to concentrate white blood cells to a portion of the whole blood sample. GLY002.1 7 Preferably, the microfluidic device also has a hybridization portion downstream of the PCR portion, the hybridization portion having an array of probes for hybridization with a predetermined target nucleic acid sequence in an amplicon from the PCR portion; And a light sensor for detecting probe hybridization in the probe array. GLY 002.1 8 Preferably, the hybridization portion has an electrode placed to receive an electrical pulse, and the probe is an electrochemiluminescence (ECL) probe for hybridization with the target nucleic acid sequence to form a probe-target hybrid. The probe-target hybridization system is configured to emit photons of light when excited by current between the electrodes. GLY002.1 9 Preferably, the light sensor is positioned to respectively register the photodiode array of the ECL probe. The easy-to-use, mass-produced and inexpensive microfluidic device accepts a biochemical sample, uses a hot lysis cell to dissolve the cells or cell cytometer, and treats the lysate. The lysis process is performed by cell extraction analysis and diagnostics in the sample, and subsequent processing and analysis of the targets are provided. Integrating this lysis unit into -224-201211533 to the device provides a simple detection step, low system component count and a simple system manufacturing process, resulting in an inexpensive inspection system. This thermal lysis process simplifies the need for detection chemistry and provides the ability to handle a wide range of species. GLY004.1 This aspect of the invention provides a test module for dissolving cells in a fluid, the test module comprising: a housing provided with a container for receiving a fluid containing cells;; a lysis reagent a reservoir containing a lysis reagent and having an outlet valve; and a lysate in fluid communication with the container for retaining the fluid during cell lysis; wherein the outlet valve is located upstream of the lysis unit and is Disposed to open when the fluid flows into the lysis section, thereby adding the lysis reagent to the fluid. GLY004.2 Preferably, the outlet valve is provided with a surface tension valve of the bore configured to form a meniscus to retain the lysis reagent in its # until the fluid contacts to remove the meniscus The lysing reagent is added to the fluid stream to enter the lysis unit. GLY004.3 Preferably, the lysis portion is configured to attract a lytic microchannel of fluid from the container by capillary action, the lysis portion having an active valve located at a downstream end of the lysis microchannel for The fluid is retained during the solubilization of the cells. GLY004.4 Preferably, the active valve is provided with a boiling start valve of a meniscus anchor, the anchor is for retaining fluid in the lysis unit, and the boiling start valve also has a valve heater. It is used to boil the liquid so that the meniscus is released from the meniscus anchor at -225 - 201211533 to resume capillary drive and flow out of the lysate. GLY004.5 Preferably, the microchannel has a cross-sectional area of the fluid from 8 square microns to 20,000 square microns. GLY004.6 Preferably, the test module also has a support substrate, wherein the lysis unit and the lysis reagent reservoir are supported by the support substrate and configured as a on-wafer laboratory (LOC) device. GLY004.7 Preferably, the test module also has a CMOS circuit between the support substrate and the lysis unit, the CMOS circuit being configured for operational control of a valve heater located at a downstream end of the microchannel. GLY004.8 Preferably, the test module also has a polymerase chain reaction (PCR) portion provided with at least one heater for heating the circulating fluid to amplify the nucleic acid sequence released from the cells. GLY004.9 Preferably, the test module also has at least one sensor, wherein the CMOS circuit is configured to use the sensor to feedback control the at least one PCR heater. GLY004.1 0 Preferably, the test module also has a plurality of heaters and at least one temperature sensor and at least one liquid sensor, so that the C Μ Ο S circuit corresponds to at least one liquid sensor To control the initial activation of the heater and to return at least one temperature sensor to control the heater power 〇GLY004.il Preferably, each of the plurality of heaters is independently operable. GLY004.1 2 Preferably, the PCR moiety is configured to have a PCR microchannel having the same cross section as the lysis microchannel of -226-201211533, the PCR portion also having an active valve located at a downstream end of the PCR microchannel to The nucleic acid sequence is used to retain the fluid during amplification. GLY004.1 3 Preferably, the test module also has a PCR mixture reservoir and a polymerase reservoir, wherein the PCR mixture reservoir comprises dNTPs, primers and buffer solutions and the polymerase reservoir comprises a polymerase. GLY004.1 4 Preferably, the test module also has an upper cover, the upper φ cover defines the lysis reagent reservoir, the PCR mixture reservoir and the polymerase reservoir, wherein the lysis unit and the PCR system Between the upper cover and the support substrate. GLY004.1 5 Preferably, the test module also has a dialysis section, wherein cells in the fluid have different sizes, and the dialysis section is configured to separate a sample larger than a predetermined valve to a portion of the sample, the section The sample is processed separately from the remaining sample containing only cells smaller than the predetermined valve. GLY004.1 6 Preferably, the nucleic acid sequence is derived from cells larger than the predetermined # valve. GLY004.1 7 Preferably, the test module also has an anticoagulant reservoir, wherein the flow system is a whole blood sample such that an anticoagulant is added to the whole blood upstream of the dialysis portion and the dialysis system is It is configured to concentrate white blood cells to a portion of the whole blood sample.

GLY004. Preferably, the test module also has a hybridization portion downstream of the PCR portion, the hybridization portion having a probe array for hybridization with a predetermined target nucleic acid sequence in the amplicon from the PCR portion; A light sensor for detecting probe hybridization in the probe array. -227- 201211533 GLY004. Preferably, the hybridization portion has an electrode placed to receive an electrical pulse, and the probe is an electrochemiluminescence (ECL) probe for hybridizing with the target nucleic acid sequence to form a probe-target hybrid. The probe-target hybridization system is configured to be excited by the current between the electrodes to emit photons of the stomach. GLY004. Preferably, the light sensor is positioned to respectively register the photodiode array of the ECL probe. The easy-to-use, mass-produced, inexpensive, and portable genetic test module accepts biochemical samples, utilizes chemical lysis subunits to lyse cells or cells, and treats the lysate. The lysis process is performed by cell extraction analysis and diagnostics in the sample, and subsequent processing and analysis of the targets are provided. Integrating this lysis unit into the module provides a simple detection step and low detection system complexity, resulting in an inexpensive detection system. GLY005. 1 This aspect of the invention provides a test module for dissolving cells in a fluid, the test module comprising: a housing having a container for receiving a sample, the sample being a fluid containing cells; in fluid communication with the container a lysate for retaining the fluid during cell lysis; and a lysis heater for heating the fluid in the lysate to dissolve the cells; wherein the test module is configured for use by capillary Acting from the container to attract fluid to the lysis unit β -228- 201211533 GLY005. Preferably, the lysis portion is configured to attract a lytic microchannel of fluid from the inlet by capillary action, the lysis portion having an active valve located at a downstream end of the lysis microchannel for use in The fluid is retained during cell lysis. GLY005. Preferably, the active valve is provided with a boiling start valve of a meniscus anchor, the anchor is for retaining fluid in the lysis unit, and the boiling start valve also has a valve heater, It is used to boil the liquid so that the meniscus is released from the meniscus anchor to recover the capillary drive and flow out of the lysate. GLY005. Preferably, the microchannel has a cross-sectional area of the fluid ranging from 1 square micron to 400 square micrometers. GLY005. Preferably, the test module also has a support substrate, wherein the lysis unit is supported by the support substrate and configured as a lab-on-lab (LOC) device. GLY005. Preferably, the test module also has a CMOS circuit between the supporting φ substrate and the lysis unit, the CMOS circuit configured to operate and control the lysis heater and located at a downstream end of the microchannel Valve heater. GLY00 5. Preferably, the test module also has a polymerase chain reaction (PCR) portion provided with at least one heater for heating the circulating fluid to amplify the nucleic acid sequence released from the cells. GLY005. Preferably, the test module also has at least one sensor, wherein the CMOS circuit is configured to use the sensor to feedback control the at least one PCR heater. -229- 201211533 GLY005. Preferably, the test module also has at least one temperature sensor and at least one liquid sensor, wherein the PCR portion has a plurality of heaters to cause the CMOS circuit to respond to at least one liquid sensor to control the The heater is initially activated and at least one temperature sensor is returned to control the heater power. GLY005. Preferably, each of the plurality of heaters is independently operable. GLY005. Il preferably, the PCR portion is configured to have a PCR microchannel having the same cross-section as the lysing microchannel, the PCR portion also having an active valve located downstream of the PCR microchannel for use during amplification of the nucleic acid sequence The fluid is retained. GLY005. Preferably, the test module also has a PCR mixture reservoir and a polymerase reservoir, wherein the PCR mixture reservoir comprises dNTPs, primers and buffer solutions and the polymerase reservoir comprises a polymerase. GLY005. Preferably, the test module also has an upper cover defining the PCR mixture reservoir and the polymerase reservoir, wherein the lysis portion and the PCR portion are interposed between the upper cover and the support substrate. GLY005. Preferably, the test module also has a dialysis section, wherein the cells in the fluid are different in size, and the dialysis section is configured to separate a sample larger than a predetermined valve to a part of the sample, and the sample of the part is The remaining samples containing only cells smaller than the predetermined valve are treated separately. GLY005. Preferably, the nucleic acid sequence is from a cell that is less than the predetermined valve. GLY005. Preferably, the test module also has an anticoagulant reservoir - 230 - 201211533, wherein the flow system is a whole blood sample such that the anticoagulant is added to the whole blood upstream of the dialysis section. GLY005. Preferably, the test module also has a hybridization portion downstream of the PCR portion, the hybridization portion having a probe array for hybridization with a predetermined target nucleic acid sequence in the amplicon from the PCR portion; A light sensor for detecting probe hybridization in the probe array. GLY005. Preferably, the hybridization portion has an electrode placed to receive an electrical φ pulse, and the probe is an electrochemiluminescence (ECL) probe for hybridizing with the target nucleic acid sequence to form a probe-target hybrid. The probe-target hybridization system is configured to emit photons of light when excited by current between the electrodes. GLY005. Preferably, the photosensor is positioned to respectively register the photodiode array of the ECL probe. The easy-to-use, mass-produced, inexpensive, and portable genetic test module accepts biochemical samples, utilizes hot lytic units to dissolve cells or cells, and treats the lysate. The lysis process is performed by cell extraction analysis and diagnostics in the sample, and subsequent processing and analysis of the targets are provided. Integrating this lysis unit into the module provides a simple detection step and low detection system complexity, resulting in an inexpensive detection system. This thermal lysis process simplifies the need for detection chemistry and provides the ability to handle a wide range of species. GD A001. 1 Aspects of the invention provide a microfluidic device for simultaneously detecting multiple states of a patient, the microfluidic device comprising: -231 - 201211533 an inlet for receiving a sample of biological material extracted from the patient a microsystem technology (MS T) layer having a detection portion having a probe array for reacting with a target molecule in the sample to form a probe-target complex, the target molecule being the medicine of the patient An indication of the state; and a light sensor for detecting the probe-target complex; wherein the probe array has more than 1,000 probes. GDA001. Preferably, the target molecule is labeled with a nucleic acid sequence and the probe array has a probe configured to hybridize to the target nucleic acid sequence to form a probe-target hybrid. GDA001. Preferably, the target molecule is a protein of interest and the probe array has a probe configured to hybridize or conjugate with the target protein to form a probe-target complex. GDA001. Preferably, the microfluidic device also has a CMOS circuit and a supporting substrate. The CMOS circuit is located between the MST layer and the supporting substrate, wherein the light sensor is inserted into the photodiode array of the CMOS circuit. GDA001. Preferably, the microfluidic device also has an array of hybridization chambers containing probes, wherein each of the probes has a nucleic acid sequence complementary to one of the target nucleic acid sequences and an electrochemiluminescence (ECL) luminophore, and each hybridization chamber An electrode having an excited state for generating the ECL luminophore, wherein the ECL luminophore emits photons of light in an excited state. GDA001. Preferably, the microfluidic device also has a nucleic acid amplification portion upstream of the hetero-232-201211533 compartment array, the nucleic acid amplification portion being configured to amplify the target nucleic acid sequence. GDA001. Preferably, the nucleic acid amplification portion is a polymerase chain reaction (PCR) portion for amplifying the target nucleic acid sequence before the target nucleic acid sequence hybridizes to the ECL probe. GDA001. Preferably, the MST layer has a plurality of MST channels configured to draw fluid containing the nucleic acid sequence of the target by capillary action through the PCR portion and into the hybridization chamber. GDA0 01. Preferably, the CMOS circuit has a pad for electrically connecting to an external device, wherein the CMOS circuit is configured to convert an output from the photodiode to show that the ECL probe hybridizes to the target nucleic acid sequence Signal and provide the signal to the pad for transmission to the external device. GDA001. Preferably, each of the probes has a functional portion that is resonated by resonance energy to quench photon emission from the ECL luminophore. # GDA001. Il preferably, the probe is configured such that when the probe forms a probe-target hybrid, the functional moiety for quenching photon emission from the ECL luminophore is further derived from the ECL luminescence group. GDA001. Preferably, the CMOS circuit is configured to provide an electrical pulse to the electrode, the electrical pulse period being less than zero. 69 seconds. GDA001. 13 Preferably, the current of the electrical pulse is between 0. 1 nai to 69. 0 nai pei. GDA001. Preferably, the CMOS circuit is configured to control the temperature of the hybridization chamber during the period of hybridization of the probe to the target nucleic acid sequence. 233- 201211533 GDA001. Preferably, the microfluidic device also has a hybrid heater. The hybrid heater is controlled by the CMOS circuit to provide thermal energy for hybridization. GDA001. Preferably, the microfluidic device also has a fluid flow path from the PCR portion to the end liquid sensor, the hybrid chamber being disposed along both sides of the fluid flow path. GDA001. Preferably, the fluid flow path is configured to draw fluid from the PCR portion to the liquid end point sensor by capillary action, and each of the hybrid chambers is configured to be filled with the fluid flow path by capillary action The fluid' is such that during use, the CMOS circuit activates the hybrid heater by reacting an output from the liquid endpoint sensor indicating that the fluid has reached the output of the liquid endpoint sensor. GDA001. Preferably, each of the hybridization chambers has a volume of less than 9,000 cubic microns. GDA001. Preferably, the distance between the photodiode and the ECL probe is less than 1,600 microns. GDA00 1. 20 Preferably, the microfluidic device also has a plurality of reagent reservoirs for treating the different reagents required for the fluid, wherein the flow system is attracted to the endpoint sensor from the inlet by capillary action without adding a source of liquid outside the microfluidic device. This easy-to-use mass-produced, inexpensive, and portable diagnostic test module accepts biological samples and processes and analyzes the samples, detecting and identifying combinations of diseases and conditions. The diagnostic capabilities of the test module provide reliability and speed for diagnosing diseases and combinations, ease of use and very low diagnostic costs. [234] DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS The present invention is directed to the main components of a molecular diagnostic system embodying an embodiment of the present invention. The full details of the system structure and operation are discussed in the following description. Referring to Figures 1, 2, 3, 104 and 105, the system has the following most important components: The test modules 1 and 1 are the same size as the conventional U S B flash drive, which can be produced very cheaply. Test modules 1 and 1 each comprise a microfluidic device, typically in the form of a lab-on-lab (LOC) device 30, which preloads reagents and typically more than one probe for molecular diagnostic analysis. (See Figures 1 and 104). The test module 10 illustrated in Figure 1 uses a fluorescent substrate detection technique to identify the target molecule, whereas the test module 1 1 of Figure 104 uses the detection technique of the electrochemiluminescence substrate. The LOC device 30 has an integrated light sensor 44 (described in detail below) for fluorescence or electrochemiluminescence detection. Test modules 10 and 11 use standard micro-USB connector 14 for power supply, data and control. Both test modules have printed circuit board (PCB) 5 7 and external power supply capacitor 3 2 and inductor 1 5 » test Modules 10 and 1 1 are for single use only, and are mass-produced and distributed for aseptic packaging for immediate use. The outer casing 13 has a large container 24 that accepts biological samples and a removable -235-201211533 sterile sealing tape 22, which preferably has a low viscosity adhesive to cover the large container prior to use. The membrane seal 408 having the membrane shield member 410 forms a portion of the outer casing 13 to reduce the humidity reduction in the test module while providing a pressure relief effect when the atmospheric pressure changes. Membrane guard 4 10 protective film seal 408 is protected from damage. The test module reader 1 2 supplies power to the test module 1 〇 or 1 1 via the micro-U S B埠1 6 . The test module reader 12 can be in many different forms, the choice of which is described later. The version of the reader 12 shown in Figures 1, 3 and 104 is an implementation of the smart phone. A block diagram of this reader 12 is shown in Figure 3. The processor 42 executes the application software from the program storage 43. The processor 42 is also interfaced with a display screen 18 and a user interface (UI) touch screen 17 and buttons 19, a cellular radio 21, a wireless network connection 23, and a satellite navigation system 25. Honeycomb radio 2 1 and wireless network connection 23 are used for communication. The satellite navigation system 25 is used to update the epidemiological database with location information. The location data can optionally be manually entered via touch screen i 7 or button 19. The data store 27 stores genetic and diagnostic information, test results, patient information, analysis and probe data for identifying the position of each probe and its array. The data storage 27 and the program storage 43 can be shared by a common device. Test module reader! The application software installed in 2 provides results analysis and other test and diagnostic information. To perform a diagnostic test, insert the test module i (or test module n) into the mini_USB port on the test module reader 12. The sterile sealing tape 22 is torn back and the biological sample (in liquid form) is loaded into the sample large container 24. Press the start button 2 to start the test via the application software. Sample stream -236-201211533 into the LOC device 30 and the device is subjected to on-board analysis to extract, culture, amplify and pre-synthesize a hybrid-reactive oligonucleotide probe with the sample nucleic acid (target) Hybrid. In the case of test module 10 (which uses detection of a fluorescent substrate), the probes are fluorescently labeled and provide the necessary excitation light by LEDs 26 mounted in the housing 13 to induce hybridized probes. Fluorescent emission (see Figures 1 and 2). In the case of test module 1 (which uses electrochemiluminescence (ECL) detection), the LOC device 30 is loaded with an ECL probe (as described above) and LED φ 26 is not required to produce luminescent emissions. In fact, the excitation current is provided by electrodes 860 and 870 (see Figure 105). The emission (fluorescence or illumination) is detected by a light sensor 44 integrated into the CMOS circuitry of each LOC device. The detection signal is amplified and converted to a digital output for analysis by the test module reader. The reader then displays the results. This information can be stored locally and/or uploaded to a network server containing patient records. The test module 10 or 1 1 is removed from the test module reader 12 and processed appropriately. # Figure 1, 3 and 104 show a test module reader 12 designed as a mobile phone/smartphone 28. Other forms of test module readers may be laptops/notebooks 101, dedicated readers 103, e-book readers 107, tablets 109 or desktop computers 105 for use in hospitals, private clinics or laboratories ( See Figure 106). The reader can interface with additional applications such as patient records, accounting, online databases and multi-user environments. It can also interface with some local or remote peripherals, such as printers and patient smart cards. Referring to FIG. 1 07, the data generated by the test module 1 can be used to update the epidemiological database and the genetic data host system stored in the epidemiological data host system 111 through the reader-237-201211533 12 and the network 125. 1 1 3 stored genetic database, electronic health record (EHR) host system 115 stored electronic health record, electronic medical record (EMR) host system 1 2 1 stored electronic medical records, and individuals Health Record (P HR) Personal health record stored by host system 123. Conversely, the epidemiological data stored in the epidemiological data host system 1 1 1 , the genetic data stored in the genetic data host system 113, and the electronic data stored in the electronic health record (EHR) host system 1 1 5 The health record, the electronic medical record stored in the electronic medical record (EMR) host system 121, and the personal health record stored in the personal health record (PHR) host system 123 can be used to access the network 1 2 5 And the reader 12 to update the digital memory in the LOC 30 of the test module 10. Referring to Figures 1, 2, 104 and 010, the reader in the mobile phone configuration uses battery power. The mobile phone reader contains all pre-loaded test and diagnostic information. Data can also be downloaded or uploaded via some wireless or contact interface to enable communication with peripheral devices, computers or online servers. The micro USB port 16 is used to connect a computer or a primary power supply for charging the battery. Figure 63 shows an embodiment of a test module 10 for a positive or negative test result requiring only a particular target, such as for testing whether a human is infected with, for example, influenza A virus H1N1. Only the USB power/indicator module 47 built for a particular purpose is required. No other readers or application software is required. The indicator 45 on the USB power/indicator module 47 alone displays a positive or negative result. This configuration is ideal for large screenings. -238- 201211533 Other items that may be provided by this system may include test tubes containing reagents for pre-treatment of specific samples and spatula and lancets for sample collection. Figure 63 shows an embodiment of a test module with a spring loaded telescopic lancet 3 90 and a lancet release button 3 92 for convenience. Satellite phones are available in remote locations. Test Module Electronics φ Figures 2 and 1 0 5 are block diagrams of the electronic components in test modules 10 and 11. The CMOS circuit integrated in the LOC device 30 has a USB device driver 36, a controller 34, a USB compatible LED driver 29, a clock 3 3, a power conditioner 3 1 , a RAM 3 8 and a program and data flash memory 40. These components provide for the entire test module 1 or 11 including the light sensor 44, temperature sensor 170, liquid sensor 174 and various heaters 152, 154, 182, 234 and associated drivers 37 and 39 and temporary storage The control of the devices 35 and 41 is 100 million. Only LED 26 (with test module 10 as an example), external power supply capacitor 32 and micro USB connector 14 are located outside of LOC device 30. LOC device 30 includes pads for connection to these external components. RAM 38 and program and data flash memory 40 have over 1,000 probe applications and diagnostic and test information (flash/security storage, such as by encryption). In the test module 1 1 designed for ECL detection, the test module 1 1 does not contain the LED 26 (see Figs. 04 and 105). The data is encrypted by the LOC device 30 for secure storage and secure communication with external devices. The LOC device 30 is loaded with electrochemiluminescent probes each having a pair of ECL excitation electrodes 860 and 870. -239- 201211533 Many types of test modules 1 are manufactured in several test formats for stock use. The difference between the different test formats is the reagents and probes for the onboard analysis. Some examples of infectious diseases that can be quickly identified using this system include: • Influenza-influenza virus A, B, C, infectious salmon virus (Isavirus), Tohogotovirus (phogotovirus) • Pneumonia-respiratory fusion Virus (RSV), adenovirus, interstitial pneumonia virus, pneumococci, staphylococcus aureus, tuberculosis, mycobacterium tuberculosis, mycobacterium bovis, mycobacteria, mycobacteria, and mycobacteria • Plasmodium falciparum, Toxoplasma gondii and other parasitic protozoa • Typhoid- typhoid bacillus • Ebola virus • Human immunodeficiency virus (HIV) • Dengue fever – Yellow fever virus • Hepatitis (A to E) • Medical source Sexual infections - such as Clostridium pneumoniae, vancomycin-resistant enterococci and drug-resistant Staphylococcus aureus • Herpes simplex virus (HSV) • Cytomegalovirus (CMV) • Epstein Barr virus (EBV) • Brain Inflammation - Japanese encephalitis virus, Zhangdipula virus • Pertussis - pertussis • Measles - paramyxovirus -240- 201211533 (Neisseria • Meningitis - Streptococcus pneumoniae and Neisseria meningi Tidis) • Anthracnose-Bacillus anthracis Examples of some genetic diseases that can be identified using this system include • Cystic fibrosis • Hemophilia • Sickle cell anemia Φ • Black Mongolian idiots • Hemochromatosis • Cerebral arterial disease • Crohn's disease • Polycystic kidney disease • Congenital heart disease • Leier's disease A few examples of cancers identified by this diagnostic system include: # • Ovarian cancer • Colon cancer • Multiple endocrine neoplasms • Retinal blastoma • Tucker Turcot Syndrome Using Nucleic Acids The above list is incomplete and the diagnostic system can be configured to detect more diverse diseases and conditions with proteomic analysis. " Detailed Structure of System Components -241 - 201211533 LOC Device The LOC device 30 is the core of this diagnostic system. The device rapidly performs the four major steps of nucleic acid substrate molecular diagnostic analysis on a microfluidic platform, i.e., sample preparation, nucleic acid extraction, nucleic acid amplification, and detection. The LOC device is also of a selective use and will be described in detail below. As discussed above, test modules 1 and 11 can take many different configurations to detect different targets. Similarly, the 'LOC device 30 can also be used to create a variety of different embodiments for the target of interest. One form of LOC device 3 0. A LOC device 310 for use in fluorescent detection of a target nucleic acid sequence of a pathogen in a whole blood sample. For the purpose of the description, the structure and operation of the LOC device 301 will now be described in detail with reference to Figs. 4 to 26 and 27 to 57. 4 is a representative diagram of the structure of the LOC device 301. For the sake of convenience, the processing stages shown in Fig. 4 are denoted by the component symbols corresponding to the functional portions of the LOC device 301 that implements the processing stage. The processing stages associated with each of the major steps in the molecular diagnostic analysis of the nucleic acid substrate are also shown: sample input and preparation 288, extraction 290, culture 291, amplification 292, and detection 2 94. The various reservoirs, chambers, valves and other components of the LOC device 301 will be described more closely below. Figure 5 is a perspective view of the LOC device 301. The device is fabricated using high volume CMOS and MST (microsystem technology) fabrication techniques. The layered structure of the LOC device 30 1 is illustrated in a schematic (not to scale) partial cross-sectional view of Figure 12. The LOC device 301 has a germanium substrate 84 that supports a COMS + MST wafer 48 that includes a CMOS circuit 86 and an MST layer 87 and has a cover 46 overlying the MST layer 87. For the purposes of this patent specification, the term "MST layer" refers to a collection of structures and layers of a sample treated with various reagents. Accordingly, these structures and components are configured to define a flow path having a feature size that will support capillary flow to drive the flow of liquid similar to the physical properties of the sample at the processing stage. In view of this, the MST layers and components are typically fabricated using surface micromachining techniques and/or stereo micromachining techniques. However, other manufacturing methods can also produce structures and components that are sized for capillary drive flow and for handling very small samples. The particular embodiment described in the specification φ shows that the MST layer is a structure and active component supported by the CMOS circuit 86, but does not have the features of the upper cover 46. However, those skilled in the art will appreciate that the MST layer does not require the underlying CMOS or even the upper cover to process the sample. The overall dimensions of the LOC device shown in the following figures are 1,760 microns X5, 8 24 microns. Of course, LOC devices made for different applications can be of different sizes. Figure 6 shows the features of the MST layer 87 that overlaps the features of the upper cover. Φ The AA to AD, AG, and AH regions shown in FIG. 6 are enlarged in FIGS. 13, 14, 3, 5, 5, 5, and 5, respectively, and are described in detail below to fully understand each of the LOC devices 301. structure. Figures 7 through 10 show the features of the upper cover 46 independently, while Figure 11 shows the structural layered structure of the CMOS + MST device 48 independently. Figures 12 and 22 illustrate the fluid interaction between the CMOS + MST device 48, the upper cover 46 & Hierarchical structure. These figures are not drawn to scale for the purpose of illustrating -243-201211533. Figure 12 is a schematic cross-sectional view through the sample inlet 68, and Figure 22 is a schematic cross-sectional view through the reservoir 54. Figure 12 clearly shows that CMOS + MST device 48 has a germanium substrate 84 that supports CMOS circuitry 86 that operates the active components within MST layer 87 thereon. The passivation layer 88 seals and protects the CMOS layer 86 from flowing into the MST layer 87. The liquid flows through both the upper cap layer 46 and the M ST channel layer 1 and the upper cap channel 94 and the MST channel 90 (see, for example, FIG. 7). And 16) » Cell transport occurs in the larger channel 94 made in the upper cover 46, while biochemical treatment takes place in the smaller MST channel 90. The size of the cell delivery channel is designed to deliver cells in the sample to a predetermined location in the MS channel 90. Cells that transport greater than 20 microns (e.g., certain white blood cells) require channel sizes greater than 20 microns, so the cross-sectional area across the flow must be greater than 400 square microns. The position of the MST channel, particularly in the LOC where no cells need to be transported, can be significantly smaller. It will be understood that the upper cover channel 94 and the MST channel 90 are generic and that the particular MST channel 90 may also be referred to as, for example, a heated microchannel or a dialysis MST channel due to its particular function. The MST channel 90 is formed by saturating the MST channel layer 100 deposited on the passivation layer 88 and patterned by a photoresist. The MST channel 90 is closed by a top layer 66 that forms the top of the CMOS + MST device 48 (shown in the figure). Although sometimes shown in separate layers, the upper cover channel layer 8 and the Benin layer 78 are formed from a single piece of material. Of course, the sheet material can also be non-unitary. Both sides of the sheet material are etched to form an upper cover channel layer 8 and a -244-201211533 reservoir layer 78, and an upper cover channel 94' is etched in the upper cover channel layer 80. The reservoirs 54, 56 are etched in the reservoir layer 78, 58, 60 and 62. Alternatively, the reservoir and the upper cover channel are formed by micromolding. Both etching and micromolding techniques are used to fabricate channels having a cross-sectional area across the fluid of up to 20,000 square microns and a minimum of 8 square microns. For various locations in the LOC device, a variety of suitable channel cross-sectional areas across the fluid can be selected. When a large number of samples or samples having a large φ composition are accommodated in the channel, a cross-sectional area of up to 20,000 square microns (e.g., a channel having a width of 200 μm in a layer having a thickness of 100 μm) is appropriate. When the channel contains a small amount of liquid or a mixture free of large cells, it is preferred that the cross-sectional area across the fluid is very small. The lower sealing layer 64 encloses the upper cover passage 94, and the upper sealing layer 82 closes the reservoirs 54, 56, 58, 60 and 62. The five reservoirs 54, 56, 58, 60 and 62 preload the reagents for the particular analysis. In the embodiments described herein, the reservoirs are preloaded with the following reagents, but can be easily replaced with other reagents: • Reservoir 54: anticoagulant, optionally including red blood cell lysate • reservoir 56: Lysis Reagents • Reservoir 58: restriction enzymes, ligases, and linkers (for linker initiation PCR (see Figure 62, excerpt from T. Stachan et al. , Human Molecular Genetics 2, Garland Science, NY and London, 1 9 9 9)) • Reservoir 60: amplification mixture (dNTPs, primers, buffers), and • reservoir 62: DNA polymerase. Upper cover 46 and CMOS + MST layer 48 are in fluid communication via corresponding openings in lower seal 64 and top -245 - 201211533 layer 66. These openings are referred to as riser 96 and descending port 92 from the MST passage 90 to the upper cover passage 94 or vice versa depending on the flow system. L Ο C device operation The operation of the LOC device 310 is described step by step as an example of pathogenic DN A in a blood sample. Of course, other types of biological or non-biological fluids can also be analyzed using appropriate reagents, test protocols, L〇C variants, and combinations or combinations of detection systems. Referring to Figure 4, the analysis of the biological sample is largely divided into five steps, including sample input and preparation 288, nucleic acid extraction 290, nucleic acid culture 291, nucleic acid amplification 292, and detection and analysis 294. The sample input and preparation steps 2 8 8 involve mixing the blood with the anticoagulant 116, followed by separation of the pathogen from the pathogen dialysis unit 70 with white blood cells and red blood cells. As best seen in Figures 7 and 12, the blood sample enters the device via sample inlet 68. Capillary action draws the blood sample along the upper lid passage 94 to the reservoir 54. - When the sample blood flow opens the surface tension valve 1 18 of the reservoir 54, the anticoagulant is released from the reservoir 54 (see Figures 15 and 22). . Anticoagulants prevent clot formation from obstructing flow. As best shown in Fig. 22, the anticoagulant agent 6 is aspirated from the reservoir 54 by capillary action and enters the MST channel 90 via the descending port 92. The lowering port 92 has a capillary initiation feature (CIF) of 1 〇 2 to control the geometry of the meniscus such that the meniscus is not fixed at the edge of the descending opening 92. The venting opening 122 in the upper sealing layer 82 allows air to replace the anticoagulant n6-246-201211533 that is aspirated from the reservoir 54. The MST channel 90 shown in Figure 22 is part of the surface tension valve 118. The anticoagulant 116 fills the surface tension valve 118 and secures the meniscus 120 to the meniscus anchor 98 of the riser 96. The meniscus 120 remains fixed to the riser 96 prior to use so that the anticoagulant does not flow into the upper cover passage 94. As blood flows through the upper cover passage 94 to the ascending port 96, the meniscus 120 is removed and the anticoagulant is drawn into the flow. 15 to 21 show an AE area which is a part of the AA φ area shown in Fig. 13. As shown in Figures 15, 16 and 17, surface tension valve 118 has three separate MST passages 90 extending between respective lower and upper risers 92, 96. The number of MST channels 90 in the surface tension valve can vary to change the flow rate of reagent into the sample mixture. When the sample mixture and reagents are mixed by diffusion, the flow rate out of the reservoir determines the concentration of the reagent in the sample stream. Thus, the surface tension valves of each reservoir are configured to meet the desired reagent concentration. The blood enters the pathogen dialysis section 70 (see Figures 4 and 15) where the target array is concentrated from the sample using a predetermined number of pore arrays 1 64. Cells smaller than the valve plaque pass through the well, while large cells cannot pass through the well. The undesired cells are either larger cells that are blocked by the well array 164 or smaller cells that pass through the well, which are transduced to the waste unit 76, whereas the target cells are still part of the assay. In the pathogen dialysis section 70 described herein, the pathogen system from the whole blood sample is concentrated for microbial DNA analysis. The array of holes is formed by a plurality of 3 micron diameter holes 1 64 that are in fluid communication with the upper cover channel 94 and input to the target channel 74. The 3 micron diameter hole 1 64 and the dialysis upwelling hole 168 in the target channel -247-201211533 74 are connected by a series of dialysis MST channels 204 (best shown in Figures 15 and 21). The pathogen is small in volume and can therefore be filled through the 3 micron diameter well 164 and filled through the target channel 74 via the dialysis MST channel 204. Cells larger than 3 microns, such as red blood cells and white blood cells, remain in the waste channel 72 of the upper cover 46, which leads to the waste receptacle 76 (see Figure 7). Other pore shapes, sizes, and aspect ratios can be utilized to isolate specific pathogens or other target cells, such as white blood cells for human DNA analysis. Detailed instructions for the dialysis section and dialysis variants are provided below. Referring again to Figures 6 and 7, fluid is drawn through target channel 74 to surface tension valve 128 of lysis reagent reservoir 56. The surface tension valve 128 has seven MST channels 90 that extend between the lysis reagent reservoir 56 and the target channel 74. When the meniscus is removed by the sample stream, the flow rate from all seven MST channels 90 will be greater than the flow rate from the anticoagulant reservoir 54, where the surface tension valve 118 of the reservoir 54 has three MST channels 9 ( It is assumed that the physical properties of the liquids are approximately equal). Thus the ratio of lytic reagent to the sample mixture is greater than the ratio of anticoagulant to the sample mixture. The lysis reagent and the target cells are mixed by diffusion in the target channel 74 in the chemical lysis unit 130. The boiling start valve 1 2 6 stops the flow until sufficient time for diffusion and lysis has occurred to release the genetic material from the target cells (see Figures 6 and 7). The structure and operation of the boiling start valve are described in detail below with reference to Figures 31 and 32. Other active valve types (as opposed to passive valves such as surface tension valve 118) have also been developed by the applicant, and other types of main-248-201211533 moving valves can be used here to replace the boiling start valve. These alternative valve designs are also described below. When the boiling start valve 126 is opened, the lysed cells flow into the mixing section 1 31 to perform restriction enzyme cleavage and linker ligation before amplification. Referring to Figure 13, when the fluid removes the meniscus of the surface tension valve 132 at the beginning of the mixing portion 131, the restriction enzyme, linker and ligase are released from the reservoir 58. This mixture flows through the length of the mixing portion 131 to be diffused and mixed. The end of the mixed φ portion 131 is a descending port 134 leading to the incubator inlet passage 133 of the culture portion 114 (see Fig. 13). The incubator inlet channel 133 feeds the mixture into a heated microchannel 210 in a crimped configuration that provides a culture chamber for holding the sample during restriction enzyme cleavage and linker ligation (see Figures 13 and 14). Figures 23, 24, 25, 26, 27, 28 and 29 show the layers of the LOC device 301 in the AB zone of Figure 6. The figures show successive layers of addition to form the structure of CMOS + MST layer 48 and upper cover 46. The AB region shows the end point of the culture unit 1 14 and the starting point of the amplification unit 1 12 . As best seen in Figures 14 and 23, the fluid fills the microchannel 210 of the culture portion 114 until it reaches the boiling start valve 1 〇 6 ' where the fluid stops to allow diffusion to occur. As discussed above, the microchannels 210 in the upstream of the boiling start valve 106 become the culture chamber containing the sample, restriction enzyme, ligase, and linker. The heater 154 is then activated and maintains a stable temperature for a specific period of time to allow for restriction enzyme cleavage and linker ligation to occur. Those skilled in the art will appreciate that this culturing step 291 (see Figure 4) is optional 'only for some nucleic acids. Required for amplification analysis type. In addition, in some examples, -249-201211533 may be required to provide a heating step at the end of the incubation period to raise the temperature above the culture temperature. The temperature rise will cause the restriction enzyme and the ligase to enter the amplification unit 1 1 2 go live. Deactivation of restriction enzymes and ligases is particularly important for the use of isothermal nucleic acid amplification. ^ After the incubation, the boiling start valve 106 is activated (opened) and the fluid continues to enter the amplification section 112. Referring to Figures 31 and 32, the mixture is filled with heated microchannels 58 in a curved configuration until reaching the boiling start valve 1〇8, which forms one or more amplification chambers. The cross-sectional view of Figure 30 clearly shows that the 'amplification mixture (dNTP, primer, buffer) is released from the reservoir 60 and the polymerase is then released from the reservoir 62 into the junction culture and amplification (114 and 112, respectively). The middle MST channel 212. Figures 35 through 51 show the layers of the LOC device 301 in the AC region of Figure 6. The figures show the layers of successive additions to form the structure of the CMOS + MST device 48 and the upper cover 46. The AC region is the end point of the amplification unit 112 and the starting point of the hybridization and detection unit 52. The cultured sample, amplification mixture, and polymerase flow through microchannel 158 to boiling start valve 108. After sufficient time of diffusion mixing, the heaters 1 54 in the microchannels 1 58 are activated for thermal cycling or isothermal amplification. The amplification mixture undergoes a predetermined number of thermal cycles or a predetermined amplification time to amplify sufficient target DNA. After the nucleic acid amplification procedure, the boiling start valve 〇8 is opened and the fluid continues to enter the hybridization and detection portion 52. The operation of the boiling start valve is described in detail below. As shown in Fig. 52, the hybridization and detection unit 52 has a hybridization chamber array 110. Figures 52, 53, 54 and 56 show in detail the hybridization chamber array 110 and a single hybridization chamber 18[deg.] at the inlet of the hybridization chamber 180 with a diffusion barrier 175'-250-201211533 which prevents target nucleic acids, probe strands and hybridization. The probes diffuse between hybridization chambers 180 during hybridization to prevent erroneous hybridization assay results. The diffusion barrier 1 75 represents a flow path of sufficient length to prevent the target sequence and probe from diffusing out of the chamber and contaminating another chamber during the time required for the probe and nucleic acid hybridization and signal to be detected, thus avoiding erroneous results . Another mechanism to prevent erroneous results is to include the same probe in multiple hybridization chambers. The CMOS circuit 86 derives a single result from the photodiode 184 corresponding to the hybridization chamber 180 φ containing the same probe. When deriving a single result, the anomalous results can be ignored or given different weights. The thermal energy required for hybridization is provided by a CMOS controlled heater 182 (described in more detail below). After activation of the heater, hybridization occurs between complementary target-probe sequences. The LED driver 29 in the CMOS circuit 86 transmits a message to the LED 26 located in the test module 10 to illuminate it. These probes only fluoresce when hybridization occurs, so the cleaning and drying steps typically required to remove unbound strands are omitted. Hybrid forced FRET probe #186 has a stem-loop structure that allows the fluorophore to emit fluorescent energy in response to the LED excitation light, as described in more detail below. Fluorescence is detected by photodiode 184 located in CMOS circuit 86 below each hybridization chamber 180 (see description of hybridization chamber below). The photodiode 184 and associated electronics for all of the hybridization chambers together form a light sensor 44 (see Figure 60). In other embodiments, the light sensor can be an array of charge coupled devices (CCD arrays). The detection signal from the photodiode 184 is amplified and converted to a digital output for analysis by the test module reader 12. Further details of the detection method are described below. -251 - 201211533 Other Detailed Description of LOC Device The modular design LOC device 301 has a number of functional components (including reagent reservoirs 54, 56, 58, 60 and 62, dialysis section 70, lysis section 130, culture section 1 14 And the expansion unit U 2), valve type, humidifier and humidity sensor. In other embodiments of the LOC device, these functional portions may be omitted, but additional functional portions may be added or the functional portions may be used for different purposes than those described above. For example, the culture portion 1 14 can be used as the first amplification portion 112 of the tandem amplification analysis system, and the chemical lysis reagent reservoir 56 is used to add the first amplification primer, dNTP, and buffer. A mixture of liquids, and a reagent reservoir 58 are used to add reverse transcriptase and/or polymerase. If the sample is subjected to chemical lysis, a chemical lysing reagent (along with the amplification mixture) may be added to the reservoir 56' or alternatively, the sample may be heated for a predetermined period of time to cause thermal dissolution in the culture section. Cell. In some embodiments, if chemical lysis is desired and it is desirable to separate the chemical lysis reagent from the primer 'dNTP and buffer mixture', an additional reservoir can be introduced upstream of the reservoir 58 adjacent to the mixture. In some cases it may be desirable to omit steps, such as incubation step 29 1 . In this case, the L0C device can be specially manufactured to avoid the reagent reservoir 58 and the culture portion 1 14 4 or the reagent can be simply not loaded in the reservoir, or if an active valve is present, the active valve is not activated to release The reagent is passed into the sample stream. Therefore, the culture portion becomes a channel for transferring only the sample from the lysis portion 1 30 to the amplification portion 112. The heaters can be operated independently, so when the reaction relies on the addition of -252-201211533 heat, such as a hot lysis reaction, setting the heater to not start during this step ensures that hot lysis does not occur in LOCs that do not require hot lysis. In the device. The dialysis unit 70 can be located at the beginning of the fluid system within the microfluidic device as shown in Figure 4 or can be located at any other location within the microfluidic device. By way of example, in some cases, it may be advantageous to dialyze after amplification stage 292 to remove cell debris prior to hybridization and detection step 294. Alternatively, two or more dialysis sections can be inserted at any location in the LOC device. Similarly, φ may be incorporated into the additional amplification portion 112 such that the multiplex target can be amplified simultaneously or sequentially before being detected by the specific nucleic acid probe in the hybridization chamber array 110. It is not necessary to perform dialysis for analyzing a sample such as whole blood, so the dialysis portion 70 is simply omitted from the sample input and preparation portion 28 of the LOC design. In some cases, it is not necessary to omit the dialysis section 70 of the LOC device, even if the analysis does not require dialysis. If the presence of the dialysis section does not cause a geometrical impediment to the analysis, the sample input and preparation section can still be used to have the LOC of the dialysis section without losing the desired function. φ In addition, the detection portion 294 can include a protein body array array that is identical to the hybrid chamber array but carries a probe that is designed to conjugate or hybridize to the sample target protein present in the non-amplified sample, rather than A nucleic acid probe designed to hybridize to a target nucleic acid sequence. It will be appreciated that the LOC devices manufactured for use with this diagnostic system are different combinations of functional components selected for a particular LOC application. The vast majority of functional units are the same for many LOC devices, and the design of additional LOC devices for new applications relates to the combination of appropriate functional components from the various functional options used in existing LOC devices. -253- 201211533 Only a few LOC devices are shown in this description. More LOC devices are illustrated in a diagram to illustrate the design flexibility of the LOC devices manufactured for this system. Those skilled in the art will readily appreciate that the LOC devices described in this specification are not exhaustive, and that many additional LOC designs are related to integrating appropriate functional combinations. Sample Type LOC Variants can accept and analyze a variety of nucleic acid or protein contents in a liquid form, including but not limited to blood and blood products, saliva, cerebrospinal fluid, urine, semen, amniotic fluid, cord blood. , breast milk, sweat, pleural effusion, tears, pericardial fluid, peritoneal fluid, environmental water samples and dip samples. Amplicon obtained from macroscopic nucleic acid amplification can also be analyzed using the LOC device; in this case, all reagent reservoirs will be empty or configured to not release their contents, and the dialysis, lysis The culture and amplification section will only be used to deliver the sample from the sample inlet 68 to the hybridization chamber 1 80 for nucleic acid detection as described above. - Some sample types require pre-treatment steps prior to input into the LOC device. For example, semen may require liquefaction and mucus may require pre-treatment with enzyme to reduce viscosity. Sample input

Referring to Figures 1 and 12, the sample is added to the large container 24 of the test module. The large container 24 is a truncated cone that is fed by capillary action into the inlet 68 of the LOC unit 301. The sample is here flowed into a 64 μηη wide X-254-201211533 60 μm deep upper cap channel 94 and is also attracted to the anticoagulant reservoir 54 by capillary action. Reagent Reservoirs A small amount of reagents required for the analysis system of a microfluidic device, such as LOC device 301, are used to allow the reagent reservoirs to contain all of the reagents required for biochemical treatment in each of the reagent reservoirs having a small volume. This volume must be less than φ 1, 〇〇〇, 〇〇〇, 〇〇〇 cubic micron, mostly less than 300,000,000 cubic micrometers, usually less than 7 〇, 00 〇, 〇 00 cubic micrometers, while the LOC device 301 is shown Less than 20,000,000 cubic microns. Dialysis Section Referring to Figures 15 through 21, 33 and 34, the pathogen dialysis section 70 is designed to concentrate pathogen target cells from the sample. As previously described, a plurality of wells 164 having a 3 micron diameter opening in the top layer 66 filter the target cells from the sample #body. As the sample flows through a 3 micron diameter well 164, the microbial pathogen enters a series of dialysis MST channels 204 through the opening and flows back into the target channel 74 via the 16 μιη dialysis upwelling 168 (see Figures 33 and 34). The remaining sample (red blood cells, etc.) remains in the upper cover channel 94. Downstream of the pathogen dialysis section 70, the upper cover passage 94 becomes a waste passage 72 to the waste reservoir 76. For a biological sample type that produces a large amount of waste, a foam insert or other porous member 49 within the outer casing 13 of the test module 10 is configured to be in fluid communication with the waste reservoir 76 (see Figure 1). The function of the pathogen dialysis section is completely dependent on the capillary work of the fluid sample -255- 201211533. A 3 micron diameter hole 164 at the upstream end of the pathogen dialysis section 70 has a capillary initiation feature (CIF) 166 (see Figure 33) to allow fluid to be drawn downward into the dialysis MST channel 204 below it. The first rising aperture 198 of the target channel 74 also has a CIF 202 (see Figure 15) to prevent fluid from easily forming a meniscus on the dialysis upwell 168. The small component dialysis section 682 shown in Fig. 71 may have a structure similar to that of the pathogen dialysis section 70. The small component dialysis section separates any small target cells or molecules from the sample by the size of the well (and shape if desired) which is adapted to allow small target cells or molecules to pass through the target channel and continue to be further analyzed. Large size cells or molecules are removed to waste receptacle 766. Thus, the LOC device 30 (see Figures 1 and 104) is not limited to pathogens having a size of less than 3 μηη, and can be used to isolate cells or molecules of any desired size. Lysis section Referring again to Figures 7, 11 and 13, the genetic material in the sample is released from the cell by chemical lysis. As described above, the lysing reagent from lysis vessel 56 is mixed with the sample stream in target channel 74 downstream of surface damper valve 128 of lysis vessel 56. However, some diagnostic assays are more suitable for use with hot lysis, or even a combination of chemical and thermal lysis of target cells. The LOC device 301 cooperates with the heating microchannel 210 of the culture portion 146. The sample stream is filled with the culture portion 1 14 and stopped at the boiling start valve 1 〇6. The culture microchannel 210 heats the sample to a temperature at which the cell membrane ruptures. In some hot lysis applications, the enzyme in the chemical lysis unit 130 is not required to be reversed, and the enzyme reaction in the chemical lysis unit 130 is completely replaced by the hot lysis. Boiling Start Valve As described above, the LOC unit 301 has three boiling start valves 126, 106 and 108. The position of these valves is shown in Figure 6. Figure 31 is an enlarged plan view showing the boiling start valve 108 shown separately at the end of the heating microchannel 158 of the amplifying portion 112. φ The sample stream 1 1 9 is drawn through the heated microchannel 158 by capillary action until reaching the boiling start valve 108. The meniscus 120 is fixed to the meniscus anchor 98 of the valve inlet 146 prior to the sample flow. The geometry of the meniscus holder 98 stops the advancing meniscus to prevent capillary flow. As shown in Figures 31 and 32, the meniscus anchor 98 is maintained by the surface tension provided by the raised opening from the MST passage 90 to the upper cover passage 94/the meniscus 120. The annular heater 152 is located around the valve inlet 146. The annular heater 152 is CMOS controlled via the boiling start valve heater contact 153. To open the valve, the CMOS circuit 86 delivers an electrical pulse to the valve heater contact 153. The ring heater 152 is heated by electrical resistance until the liquid sample 119 is boiled. This boiling causes the meniscus 120 to be removed from the valve inlet 146 and begin to wet the upper cover passage 94. Once the upper cover passage 94' is wetted, the capillary action is restored. The fluid sample 112 is filled with the upper lid passage 94 and flows through the valve lowering port 150 to the valve outlet 148 where the capillary action driven liquid flow continues along the amplifying portion outlet passage 160 to the hybridization and detection portion 52. The liquid sensor 1 74 is placed before and after the valve for diagnosis -257- 201211533 It will be understood that once the boiling start valve is opened, it can no longer be closed. However, since the LOC device 301 and the test module 1 are single-use devices, there is no need to close the valve. Culture section and nucleic acid amplification section The culture section 114 and the amplification section 112 are shown in Figs. 6, 7, 13, 14, 23, 24, 25, 35 to 45, 50, and 51. The culture portion 114 has a single strip of heated culture microchannels 210 which are etched into a meandering pattern in the MST channel layer 1〇〇, starting at the lowering port 134 and finally boiling the activation valve 1〇6 (see FIGS. 13 and 14). . Controlling the temperature of the culture section 1 14 allows the enzyme reaction to occur with higher efficiency. Similarly, the amplifying portion 12 has a heated amplifying microchannel 158 (see Figs. 6 and 14) that begins in a curved structure with the boiling start valve 106 leading to the boiling start valve 108. When mixing, culture, and nucleic acid amplification occurs, the valves stop the flow to retain the target cells in the heated culture or amplification microchannel 210 or 158. The curved pattern of the microchannels also promotes (to some extent) the mixing of the target cells with the reagents. In the culture unit 4 and the amplification unit 1 1 2, the sample cells and reagents are heated by a heater 1 54 controlled by a pulse width modulation (PWM) CMOS circuit 86. Each curved corner of the heated culture microchannel 210 and the expanded microchannel 158 has three independently operable heaters 154 extending between the individual heater contacts 156 (see Figure 14)), which provides two-dimensional control of the input heat flux density. As best shown in Figure 51, the heater 154 is supported by the top layer 66 and embedded in the lower layer - 258 - 201211533. The material of the heater is TiAl, but many other conductive metals are also suitable. The elongated heater 154 is parallel to the longitudinal length of each of the channel portions forming the wide bend of the curved structure. In the amplifying portion 112, each wide curve can be controlled via a separate heater as a separate P C R chamber. The use of a small volume of amplicons required for the analysis system of the microfluidic device, such as LOC device 301, allows for the use of a small volume of amplification mixture in amplification portion 112 for amplification. This volume must be less than 400 nanoliters, mostly less than φ 17 〇 liters, usually less than 70 liters, and the LOC device 301 is exemplified by 2 nanoliters to 30 nanoliters. Increased heating rate and better diffusion mixing The small cross-sectional area of each channel section increases the heating rate of the amplification fluid mixture. The distance between all fluids and heater 154 is quite short. Reducing the cross-sectional area of the channel (i.e., the cross section of the augmented microchannel 158) to less than 1 〇〇, the squared micrometer can achieve a significantly higher heating rate than the "large scale" device. Photolithography • Manufacturing techniques such that the amplifying microchannel 158 has a cross-sectional area of less than 1 6,000 square micrometers across the flow path, which provides a substantially higher heating rate. Dimensional features of 1 micron are easily obtained using lithography manufacturing techniques. If only a very small amount of amplicons are required (in the case of LOC device 301), the cross-sectional area can be reduced to less than 2,500 square microns. A traversing fluid between 400 square microns and 1 square micrometer for a diagnostic analysis performed on a LOC device with 1,000 to 2,000 probes and requiring "input sample to get results" within one minute The cross-sectional area is appropriate. The heater element in the amplification microchannel 158 heats the nucleic acid sequence at a rate of greater than 80 -259 - 201211533 per second to temperature (K) in most cases, at a rate of greater than 100 Torr per second. Typically, the heater element heats the nucleic acid sequence at a rate greater than 1,000 psi per second, and the heater element often heats the nucleic acid sequence at a rate greater than 10,000 Torr per second. Typically, depending on the requirements of the analytical system, the heater element is greater than 100,000 sec per second, greater than 1,000,000 sec per second, greater than 10,000,000 sec per second, greater than 20,000,000 sec per second, and greater than 40,000,000 per second. The nucleic acid sequence is heated at a rate of greater than 80,000,000 每秒 per second and greater than 160,000,000 每秒 per second. The small cross-sectional area channel is also beneficial for the diffusive mixing of any reagent with the sample fluid. The phenomenon of one liquid diffusing to another liquid is most pronounced near the interface between the two liquids before diffusion mixing is completed. The concentration decreases as the distance from the interface increases. The use of microchannels having a relatively small cross-sectional area across the fluid direction causes the two fluids to flow closer to the interface for faster diffusion mixing. Reducing the cross-sectional area of the channel to less than 100,000 square microns provides a significantly higher mixing rate than "large scale" equipment. The lithography manufacturing technique allows the microchannel to cross the flow path with a cross-sectional area of less than 1 6,000 square microns, which provides a significantly higher mixing rate. If only a very small volume (in terms of LOC device 301) is required, the cross-sectional area can be reduced to less than 2,500 square microns. 'Between 400 cm and 1 square micron for diagnostic analysis on a LOC device with 1, 〇〇〇 to 2,000 probes and requiring "input sample, result" in 1 minute The cross-sectional area across the fluid is appropriate. Rapid Thermal Cycle Time -260- 201211533 Keep the sample mixture close to the heater and use a very small amount of fluid to perform a rapid thermal cycle during nucleic acid amplification. For a target sequence of up to 150 base pairs (bp) in length, each thermal cycle (i.e., denaturation, adhesion, and extension of the primer) is completed in less than 30 seconds. In most diagnostic analyses, the individual thermal cycle time is less than 11 seconds and most are less than 4 seconds. For LOC device 30, which performs some of the most common diagnostic assays, the thermal cycle time for a target sequence of up to 150 base pairs (bp) is between 0.45 seconds Φ and 1.5 seconds. This rate of thermal cycling allows the test module to complete the nucleic acid amplification process in as little as 10 minutes; usually less than 220 seconds. For most analyses, the amplification produces enough amplicons within 80 seconds of the sample fluid entering the sample inlet. Many analyses produce enough amplicons in 30 seconds. When a predetermined number of amplification cycles are completed, the amplicon is fed to the hybridization and detection portion 52 via the boiling start valve 108. • Hybridization Chambers Figures 52, 53, 54, 56 and 57 show hybridization chambers 180 in the hybridization chamber array Π0. The hybridization and detection section 52 has a 24x45 array 1 10 of hybrid chambers 180 each having a hybrid-reactive FRET probe 186, a heater element 182, and an integrated photodiode 184. The photodiode 1 84 is introgressed to detect fluorescence produced by hybridization of the target nucleic acid sequence or protein to the FRET probe 186. Each photodiode 184 is independently controlled by a CMOS circuit 86. Any substance between the FRET probe 186 and the photodiode 1 84 must be transparent to the emitted light. Therefore, the wall portion 97 between the probe 186 and the photodiode 184 can also be optically transmitted by the emitted light at -261 - 201211533. In LOC device 301, wall portion 97 is a thin layer of hafnium oxide (about 0.5 microns). Direct incorporation of photodiode 184 beneath each hybridization chamber 180 allows for a detectable fluorescent signal to be produced by the extremely small probe-target hybrid volume (see Figure 54). This small amount allows the use of a small volume of hybridization chamber. The amount of detectable probe-target hybrid required for pre-hybridization probes must be less than 270 picograms (corresponding to 900,000 cubic micrometers), in most cases less than 60 picograms (corresponding to 2〇) 〇, 〇〇〇 cubic micron), typically less than 12 picograms (corresponding to 40,000 cubic micrometers), and is less than 2.7 picograms (corresponding to a chamber volume of 9,000 cubic micrometers), as exemplified by the LOC device 301 shown in the drawings. Of course, reducing the size of the hybridization chamber allows for a higher density chamber, thus allowing the LOC device to have more probes. In LOC unit 301, the hybrid has more than 1,000 chambers (i.e., less than 2,250 square microns per chamber) in an area of 1,500 microns by 1,500 microns. The smaller volume also reduces reaction time, so hybridization and detection can be faster. Another advantage of requiring a small number of probes in each chamber is that only a very small amount of probe solution needs to be spotted during manufacture of the LOC device. In the room. Embodiments of the LOC device of the present invention can be spotted using a probe solution having a volume of 1 liter or less. After the nucleic acid amplification, the boiling start valve 1〇8 is activated and the amplicon flows along the flow path 176 and flows into each of the hybridization chambers 180 (see FIGS. 52 and 56). The end point liquid sensor 187 shows hybridization. The chamber 180 is filled with the amplicon and the point at which the heater can be activated. -262- 201211533 After a sufficient hybridization time, LED 26 (see Figure 2) is actuated by opening the openings in each of the hybridization chambers 180 to provide a light window 136 to expose the FRET probe 186 to the excitation radiation (see Figures 52, 54 and 56). . The LED 26 is illuminated for a time sufficient to induce a high intensity fluorescent signal from the probe. The photodiode 184 is shorted during excitation. The preprogrammed delay 300 (see, after the reference, the photodiode 184 is enabled and detected in the presence of no excitation light. The active Φ 185 at the photodiode 1 84 (see Figure 54) The incident light is converted into a photocurrent that can then be measured using a CMOS circuit. The hybridization chambers 180 each carry a probe for detecting a single target nucleic acid sequence, and each hybridization chamber 180 can carry more than 1,000 detections if desired. Probes of different targets. Alternatively, many or all of the hybridization chambers may carry the same probe to repeatedly detect the same target nucleic acid. In this manner, the preparation of the probe in the hybridization chamber array 110 results in a credible result. If the degree is increased, if it is desired to provide a single result by entanglement of the photodiodes adjacent to the hybridization chambers, the skilled artisan will appreciate that there may be more than 1,000 different probes on the hybridization chamber array 110. The needle is determined according to the analytical specification. Hybridization chambers for electrochemiluminescence detection Figures VII, 120, 138 and 139 show the hybridization chamber 180 used in the ECL variant L0C variant L 729 of the LOC device. In the implementation, the 24x45 array of hybridization chambers Columns 1 10 each have a cross-reactive ECL probe 2 3 7 that is in registration with a corresponding array of photodiodes 184 integrated into CMOS. It is configured for use with a fluorescent needle length of 9 2) 86 〇 The weighted composite enthalpy to the variable-packaged light-263-201211533 is similar to the LOC device detected, and each photodiode 184 is intrusive for detecting the hybridization of the target nucleic acid sequence or protein with the ECL probe 23 7 ECL. Each of the photodiodes 184 is independently controlled by a CMOS circuit 86. Similarly, the transparent wall portion 97 between the probe 186 and the photodiode 1 84 can be transmitted by the emitted light. Photodiode 184 in close proximity to each hybridization chamber 180 allows the very small amount of probe-target hybrid to still produce a detectable ECL signal (see Figure 97). This small amount allows the use of a small volume of hybridization chamber. The detectable amount of the probe-target hybrid requires a probe amount prior to hybridization of less than 270 picograms (corresponding to a chamber volume of 900,000 cubic micrometers), and in most cases less than 6 picograms (corresponding to 2 00,000 cubic micrometers, typically less than 12 picograms (corresponding to 40,000 cubic micrometers), and is less than 2.7 picograms (corresponding to a chamber volume of 9,000 cubic micrometers) as an example of a LOC device as shown. Of course, reducing the size of the hybrid chamber allows for a higher density chamber, thus allowing the LOC device to have more probes. In the LOC device shown, the hybrid has more than one, one chamber (i.e., less than 2,250 square microns per chamber) in an area of 1,500 microns by 1,500 microns. Smaller volumes also reduce reaction time, so hybridization and detection can be faster. Another advantage of requiring a small number of probes in each chamber is that only a very small amount of probe solution needs to be spotted into each chamber during manufacture of the LOC device. Taking the LOC device in the illustrated example as an example, the required amount of probe can be spotted with a solution volume of 1 picoliter or less. After nucleic acid amplification, the boiling start valve 108 is activated and the amplicon flows along the flow path 76 and flows into each of the hybridization chambers 180 (see Figures 52 and 139). The end liquid sensor 178 shows the The hybridization chamber 180 is filled with the -264-201211533 point of the amplicon so that the heater 108 can be activated. The photodiode 184 is enabled to collect the ECL signal after sufficient hybridization time. The ECL excitation driver 39 (see Figure 105) then activates the ECL electrodes 860 and 870 for a predetermined period of time. The photodiode 184 maintains a short period of activity after the ECL excitation current is stopped to maximize the signal to noise ratio. For example, if the photodiode 1 84 remains active for 5 times the decay lifetime of the luminescent emission, the signal will decay to a small φ of 1% of the initial enthalpy. Light incident on the photodiode 184 is converted into a photocurrent, which can then be measured using a CMOS circuit 86. Protein Body Detection Chambers Some LOC variants, such as the LOC variant L 729, are configured to perform homogeneous protein detection on crude cell lysates in a protein body detection chamber array (see, eg, Figures 1216 to 124.3 of Figures 16 and 120) for detection. Host cells and/or pathogenic proteins. The production and configuration of the protein body detection chamber arrays 1 24.1 to Φ 124.3 are identical to the hybrid chamber arrays (see Figures 52, 53, 54, and 56). Each protein body detection chamber has a diffusion barrier 175 at the inlet to prevent diffusion of the sample and reagents between the chambers, thus avoiding erroneous results (see Figures 84 and 85, which are in the DC and DD regions of Figure 81). When protein hybridization or conjugation is desired, thermal energy is provided by CMOS controlled heaters 182 in each chamber. In some embodiments, the endpoint liquid sensor 1 78 is used to display the point at which the protein body detection chamber is full of samples so that the heater 1 82 can be activated. After sufficient time, the fluorescent or electrochemiluminescent signal generated after protein recognition is detected by the light sensor 44 -265 - 201211533

Humidifier and Humidity Sensor The AG zone of Figure 6 indicates the location of the humidifier 1 96. The humidifier prevents the reagents and probes from evaporating during operation of the LOC device 301. Preferably, as shown in the enlarged view of Figure 55, the reservoir 188 is in fluid communication with the three evaporators 1 90. The water reservoir 188 holds molecular biological grade water and is sealed during manufacture. As best seen in Figures 55 and 61, water is drawn by capillary action to the three lower rise ports 194 and along the individual water supply passages 192 to the three riser ports 193 of the evaporator 190. The meniscus is fixed to each of the risers 193 to retain water. The evaporator has a ring heater 191 that surrounds the riser 193. The ring heater 191 is connected to the top metal layer 195 of the CMOS circuit 86 by a conductive post 376 (see Figure 37). At startup, the ring heater 191 heats the water to evaporate and wet the surrounding devices. Figure 6 also shows the position of the humidity sensor 23 2 . However, as best shown in the enlarged view of the AH zone of Fig. 58, the humidity sensor has a capacitive comb structure. The lithographically etched first electrode 296 and the lithographically etched second electrode 298 are opposed to each other such that their teeth are interleaved. The opposing electrode forms a capacitor having a capacitance that can be monitored by CMOS circuit 86. As the humidity increases, the permittivity of the air gap between the electrodes increases, causing the capacitance to increase. It is important that the humidity sensor 23 2 is adjacent to the humidity of the hybridization chamber array 1 10 ' to slow the evaporation of the solution containing the exposed probe. Feedback Sensor -266- 201211533 Temperature and liquid sensors are injected into the LOC device for feedback and diagnosis during device operation. Referring to Fig. 35, 170 is distributed to the entire portion of the amplifying portion 112. Similarly, there are nine temperature sensors 170. Each of the sensors causes a pole junction transistor (BJT) to monitor the fluid temperature and the circuit 86 » the CMOS circuit 86 utilizes this for accurate thermal cycling and thermal lysis and incubation in the hybrid chamber 180, The CMOS circuit 86 makes the month a temperature sensor (see Fig. 56). The hybridization heater is temperature dependent and the CMOS circuit 86 utilizes the 180 temperature reading. The LOC device 301 also has a number of MST 174 and an upper cover channel liquid sensor 208. Figure 35 shows a row of MST 174 at one of the other corners of 158. Preferably, as shown in FIG. 37, the MST channel liquid is opposed to one of the top metal layers 195 of the CMOS structure 86. The liquid closes the current between the electrodes to the position of the finger. Figure 25 shows that the upper cover channel liquid sensor 208 is deposited between the I electrodes 218 and 220 relative to the TiAl electrode pairs 218 and 220 as a gap 222 in which the circuit is held open. CMOS circuit 86 utilizes this feedback to monitor flow when liquid is present. 3 0 1 everywhere to provide nine temperature sensors. The culture unit 1 1 4 also uses a 2x2 array of doubles for feedback to CMOS to control nucleic acid amplification. 3 The resistance temperature of the hybrid heater 182 182 is derived to derive the hybrid chamber channel liquid sensor shown in the heated microchannel channel liquid sensor body sensor 174 exposed area to form an enlarged perspective view of the presence of the sensing . On the page layer 66. In the absence of liquid i shut the circuit and the -267- 201211533 non-gravity dependent test module 1 is non-directional dependent. These modules do not need to be fixed to a smooth surface to operate. The fluid flow driven by the capillary action and the external tubing that does not have to be connected to the auxiliary device make the module truly portable and can be easily inserted into a similar portable handheld reader, such as a mobile phone. Operation with non-gravity dependence means that these test modules also contribute to all practical applications. They can withstand shock and vibration, they can operate on moving vehicles, or when the mobile phone is moving. Dialysis variants The dialysis design of the white blood cell marker LOC device 301 as described above is labeled with the pathogen. Figure 5 is a schematic cross-sectional view of a dialysis section 3 2 8 designed to concentrate white blood cells from a blood sample for human DNA analysis. It will be appreciated that the structure is substantially identical in construction to the dialysis section 70 of the above-described pathogen, except that a hole in the form of a 7.5 micron diameter hole 165 restricts the passage of white blood cells from the upper cap passage 94 to the dialysis MST channel 204. In the case where the sample is analyzed and the presence of hemoglobin from the red blood cells interferes with the subsequent reaction step, the addition of red blood cell lysis buffer with the anticoagulant (see Figure 22) in reservoir 54 will ensure that most of the The dissolved red blood cells (and therefore the hemoglobin) will be removed from the sample during this dialysis step. Frequently used red blood cell lysis buffers are 0.15 M NH4CL, 10 mM KHC03, O.lmM EDTA pH 7.2-7.4, but those skilled in the art will appreciate that any buffer that effectively dissolves red blood cells can be used. -268- 201211533 At the downstream of the leukocyte dialysis section 328, the capping channel 94 becomes the target channel 74 to continue the leukocytes as part of the detection. Also in this case, the dialysis riser 1 68 leads to the waste channel 72 to remove all of the smaller cells and components in the sample. It should be noted that this dialysis variant only reduces the concentration of the undesired sample in the target channel 74. Figure 72 schematically illustrates a large group of φ divided dialysis portions 68 6 that also separate any large components from the sample. The pores in the dialysis section are custom-made such that their size and shape retain the component of the subject of interest in the target channel for further analysis. When using the leukocyte dialysis section described above, most, but not all, of the smaller cells, organisms or molecules flow to the waste reservoir 768. Therefore, other embodiments of the LOC device are not limited to separation sizes greater than 7.5. A white blood cell of μπι, which can also be used to isolate cells, organisms or molecules of any desired size. # Dialysis section having a flow passage to prevent trapping of air bubbles The following describes an embodiment of the LOC apparatus called LOC variant VIII 518, as shown in Figs. 65, 66, 67 and 68. This LOC device has a dialysis section that is filled with a fluid sample and that no air bubbles are trapped in the channel. LOC Variant VIII 518 also has other layers of material known as interface layer 594. The interface layer 594 is between the upper cap channel layer 80 and the MST channel layer 100 of the CMOS + MST device 48. The interfacial layer 594 allows for more complex fluid intercommunication between the reagent reservoirs and the MST layer 87 without increasing the size of the tantalum substrate 84. -269- 201211533 Referring to Figure 66, the bypass channel 600 is designed to introduce a time delay as the fluid sample flows from the interface waste channel 604 to the channel 602 of the interface. This time delay allows the fluid sample to flow through the dialysis MST channel 204 to the dialysis riser 168 where a meniscus is formed. The sample fluid is filled from the point upstream of the dialysis riser 168 of all of the dialysis MST channels 204 by the capillary initiation feature (CIF) 202 at the riser of the channel 602 from the bypass channel 600. 〇 2. Without the bypass channel 600, the channel 602 of the interface still begins to charge from the upstream end, but finally the advancing meniscus reaches and rises beyond the rising mouth of the MST channel that is not full, resulting in the formation of bubbles there. . The trapped air reduces the rate at which the sample flows through the leukocyte dialysis unit 3 28 .

Nucleic Acid Amplification Variants Direct PCR Conventionally, P CR requires extensive purification of target DNA prior to preparation of the reaction mixture. However, with appropriate changes in chemical properties and sample concentrations, it is possible to perform nucleic acid amplification or direct amplification with minimal DNA purification. When the nucleic acid amplification program is PCR, this method is called direct PCR. When the nucleic acid amplification is carried out in a controlled constant temperature LOC unit, the method is directly thermostated. Direct nucleic acid amplification techniques have significant advantages when used in LOC devices, especially with regard to simplifying the fluid design required. Adjustments to the amplification chemistry of direct PCR or direct isothermal amplification include increasing buffer strength, using highly active and pr〇cessivity polymerases and additions to potential polymerase inhibitors. It is also important to dilute the inhibitor in the sample -270- 201211533. In order to utilize direct nucleic acid amplification techniques, the LOC device has two additional features. The first feature is a reagent reservoir (eg, imager 58) that is suitably sized to supply a sufficient amount of the extender or diluent to minimize the final concentration of the sample that may interfere with the amplification chemistry. Allows for successful nucleic acid amplification. The desired dilution factor of the non-fine component is between 5 and 20 times. When using different LOC structures to confirm the maintenance of a sufficiently high target nucleic acid for amplification and detection, such as in the pathogen dialysis section of Figure 4 (further illustrated in Figure 6), in sample extraction The dialysis section is used to effectively concentrate the pathogens that are small enough to enter the amplification section 292 and discharge the larger cells to the waste container 76. In the aspect, the dialysis section is used to selectively remove the eggs in the plasma while retaining the cells of interest. A second LOC structural feature that supports direct nucleic acid amplification • a broad ratio design to adjust between the sample and the amplified mixed component, for example, to ensure that the inhibitor associated with the sample is diluted to a single step In the preferred range of 5 to 20 times, the length and cross section of the channel are designed such that the sample channel at the mixing start has a flow impedance four to nine times higher than the channel through which the reagent mixture flows. . The flow impedance of the microchannel is controlled via control design geometry. In terms of a fixed cross-sectional area, the micro-impedance increases linearly with the length of the channel. For hybrid designs, the flow impedance in the microchannel is most dependent on the minimum cross-sectional area. The most cytosolic sample of the mixed reaction component in the design of the intrusion 8 will then have a sequence concentration. In this case, the concentration of the upstream body is 290. The other implementation of white matter and salt is the channel deep mixing ratio. A mixed step and reagent flow upstream of the flow impedance can easily flow through the channel. Important is the size. Example -271 - 201211533 For example, when the aspect ratio is extremely non-uniform, the flow resistance of a microchannel with a square cross section is inversely proportional to the cube of the smallest vertical dimension. Reverse Transcriptase PCR (RT-PCR) When the sample nucleic acid species analyzed or extracted are RNA, such as from RNA viruses or messenger RNAs, the RNA must first be reverse transcribed into complementary DNA (cDNA) before PCR amplification. The reverse transcription reaction can be carried out in the same chamber as the PCR (one-step RT-PCR), or it can be an initial reaction (two-step RT-PCR). In the LOC variants described herein, a one-step RT-PCR can be performed by simply adding a reverse transcriptase to a reagent reservoir 62 containing the polymerase and staging the heater 1 54 to perform the reverse transcription step. The nucleic acid amplification step is carried out. The two-step RT-PCR can also perform the reverse transcription step by using the reagent reservoir 58 to store and distribute the buffer, the primer, the dNTP, and the reverse transcriptase, and then perform the reverse transcription step in the amplification portion 112. Amplification in the usual manner is simply accomplished. Constant Temperature Nucleic Acid Amplification For some applications, thermostatic nucleic acid amplification is a preferred method of nucleic acid amplification, so that the reaction component does not need to be subjected to repeated cycles of different temperatures, but the amplification is maintained at about 3 7 °C to 4 1 °C at constant temperature. Some thermostated nucleic acid amplification methods have been described as 'including strand-substituted amplification (SDA), transcription-mediated amplification (TMA), nucleic acid sequence basal amplification (NASBA), recombinase polymerase amplification (RPA), unwinding Enzyme-dependent thermostated DNA amplification -272-201211533 (HDA), rolling-cycle amplification (RCA), branched-type amplification (ram), and circular mediation-enhanced amplification (LAMP), any of these methods or others The isothermal amplification method can be used in a particular embodiment of the LOC device described herein. In order to perform a thermostatic nucleic acid amplification, the reagent reservoirs 60 and 62 adjacent to the amplification section will carry appropriate reagents for a particular constant temperature method rather than carrying a PCR amplification mixture and a polymerase. For example, in the case of SDA, the reagent sump 60 contains amplification buffer, primers, and dNTPs, and the reagent reservoir 62 contains appropriate endonucleases and exo-DN A polymerase. In the case of RP A, the reagent reservoir 60 contains amplification buffer, primer, dNTP and recombinase protein. The reagent reservoir 62 contains a strand-substituted DNA polymerase such as Bsu. Similarly, in the case of HD A, the reagent reservoir 60 contains amplification buffer, primers and dNTPs, which contain appropriate DNA polymerase and helicase to unwind the double strand DNA instead of using heat. Those skilled in the art will appreciate that the necessary reagents can be dispensed into the two reagent reservoirs in any manner suitable for the nucleic acid amplification method. To amplify a viral nucleic acid from an RN A virus such as HIV or hepatitis C virus, NASBA or TMA is appropriate because it does not require transcription of the RNA to cdNA first. In this example, the reagent reservoir 60 is filled with an expansion buffer, an initiator, and dNTP, and the reagent reservoir 62 is filled with an RNA polymerase, a reverse transcriptase, and an arbitrary RNase. Some forms of thermostatic nucleic acid amplification must have an initial denaturation cycle to separate the double stranded DNA template before maintaining the temperature suitable for constant temperature nucleic acid amplification for the reaction to proceed. This can be easily achieved in all of the LOC devices described herein - 273 - 201211533, since the temperature of the mixing in the amplification section 112 can be carefully controlled by the heater 154 in the amplification microchannel 158. (See Figure 14). Thermostatic nucleic acid amplification is more tolerant to potential inhibitors in the sample and is therefore generally suitable for situations where direct nucleic acid amplification from the sample is desired. Therefore, thermostatic nucleic acid amplification is sometimes used for LOC variant XLIII 673, LOC variant XLIV 674 and LOC variant XL VII 677, etc., respectively, shown in Figures 73, 74 and 75. Direct thermostatic amplification can also be combined with one or more of the pre-amplification dialysis steps 70, 686 or 682 as shown in Figures 73 and 75, and/or the pre-hybridization dialysis step 682 as shown in Figure 74, respectively. Part of the target cells in the sample are concentrated prior to nucleic acid amplification, or unwanted cell debris is removed before the sample enters the array of hybridization chambers. Those skilled in the art will appreciate that any combination of pre-amplification dialysis and pre-hybridization dialysis can be used. Thermostatic nucleic acid amplification can also be performed in parallel amplification sections such as Figures 64, 69 and illustrated, multiplex and some Methods of thermostatic nucleic acid amplification such as the LAMP line are compatible with the initial reverse transcription step to amplify RNA. Other design variant flow rate sensors In addition to the temperature and liquid sensors, the LOC device can also be equipped with a C Μ 0 S-controlled flow rate sensor 704, as shown in Figure 9.4 and L Ο C variants. Graphical illustration in X 72 8 (see Figures 76 to 92). These sensors are used to measure the flow rate in two steps. In the first step, the temperature of the bent heater element 8 1 4 - 274 - 201211533 is determined by applying a low current and measuring the voltage to determine the resistance of the bent heater element 8 14 , so the element 8 The temperature of 14 is calculated using the known relationship between the resistance of the heater element and the temperature. In this stage, very little heat subsides in the element 814, and the temperature of the liquid in the channel is equal to the calculated temperature of the element 814. In the second step, a higher current is applied to the bent heater element 8 1 4 to increase the temperature of the element 8 14 and some of the heat is lost to the flowing liquid. By again measuring the voltage of the element 8 14 when φ is applied with a higher current, the new resistance of the element 8 14 is measured and the increased temperature is again calculated by the CMOS circuit 86. The flow rate of the liquid is determined by the new temperature of the bent heater element 814 and the known temperature of the sample liquid calculated in the first step. From the known channel cross-section geometry and the flow rate, the flow rate of the liquid in the channel is calculated. Protein Detection Variants # Some embodiments of this LOC device use homogeneous protein detection to detect specific proteins in the crude cell lysate. A variety of homogeneous protein detection assays for use in the implementation of this LOC device have been developed. Typically, these assays utilize antibodies or aptamers to capture the target protein. In one type of assay, aptamer 141 bound to a particular protein 142 is labeled with two different fluorophores or luminescent groups 143 and 144, The function is as a donor and recipient in fluorescence resonance energy transfer (FRET) or electrochemiluminescence resonance energy transfer (ERET) reactions (see Figure 〇 8〇 and -275- 201211533 108B) » Donor M3 and recipients 144 is both linked to the same aptamer 141, and the separation alteration is caused by a change in configuration when the target protein 142 is bound. For example, in the absence of the target, the aptamer 141 forms a configuration in which the donor is in close proximity to the recipient (see Figure 108A); when combined with the target, the new configuration results in the donor and acceptance Large separation between the two (see Figure 108B). When the recipient is a quencher and the donor is a luminescent group, the effect of combining with the target is an increase in light emission of 250 or 8 62 (see Figure 10 8 B). The second type of assay utilizes two antibodies 145 or two aptamers 141 (see Figures 109A, 109B, 110A and 110B) that must bind independently of the target protein 142, non-overlapping epitopes or regions. These antibodies 145 or aptamer 141 are labeled by different fluorophores or luminescent groups 143 and I44, and their functions are donated in fluorescence resonance energy transfer (FRET) or electrochemiluminescence resonance energy transfer (ERET) reactions. And recipients. The fluorophore or luminophore 143 and M4 form part of a pair of short complementary oligonucleotides 147 that are linked to the antibodies or aptamers via a long, bendable linker 149 ( See Figures 109A and 110A). Once the antibodies 145 or aptamers 141 bind to the target protein M2, the complementary oligonucleotides 147 find each other and hybridize to each other (see Figures 109B and 10B). This allows the donors and recipients 143 and 144 to be in close proximity to each other 'causing effective FRET 25〇 or ERET 862 as a signal for target protein detection » in order to ensure no or little background signal (because with the two antibodies 1 45 or aptamer 141-linked oligonucleotides are caused by hybridization to each other when they are not bound to the protein 142-276-201211533), the length and sequence of the complementary oligonucleotides 147 must be carefully selected so that The dissociation constant (kd) of the double strands is relatively high (~5 μΜ). Thus, when the free antibody or system indicated by these oligonucleotides is mixed at a concentration of nanomolar (far less than the concentration of kd), the possibility of double-strand formation and the resulting FRET 250 or ERET 8 62 signals can be ignored. However, when two antibodies 145 or two aptamers 141 are bound to the target protein 142, the local concentration of the oligonucleotide 147 will be much higher than φ, resulting in almost complete hybridization and a detectable FRET. 250 or ERET 862 signal. The choice of fluorophores and luminophores is an important consideration when designing homogeneous protein assays. The crude cell lysate is usually turbid and may contain substances with autofluorescence. In such cases, donor-recipient pairs 143 and 144 with long-term fluorescence or electrochemiluminescence and optimized to produce maximum FRET 250 or ERET 8 62 are desired. One such donor-recipient pair of ruthenium chelate and Cy 5, which has previously been shown to significantly increase the signal-to-background ratio of the system compared to other donor-recipient pairs, by allowing interference The background is read after the fluorescent, electrochemiluminescence or scattered light has decayed. The ruthenium chelate and AlexaFluor 647 or ruthenium chelate and fluorescein FRET or ERET pair also have good effects. The sensitivity and specificity of this method is similar to enzyme-linked immunosorbent assay (ELISA), but does not require sample processing. In some embodiments of the LOC device, one of the antibodies M5 or one of the aptamers 141 is linked to the bottom of the protein body detection chamber 124 (see, eg, Figures 1 16 and 120), And the protein lysate system -277-201211533 is combined with the other antibody 145 aptamer 141 during the dissolution in the chemical lysis unit 130 to promote the first step before entering the protein body detection chamber 丨24. A combination of antibody 145 or aptamer 141. This increases the rate at which subsequent detectable signals are generated, since only one conjugate or hybrid event is required within the proteomic detection chamber. Photodiode Figure 54 shows a photodiode 184 that is shunted into the LOC device 301 CMOS circuit 86. The photodiode 184 is part of the CMOS circuit 86 without additional masking or smearing. This is a significant advantage of CMOS photodiodes over CCDs, an alternative to the sensing technique that can be integrated on the same wafer or fabricated on adjacent wafers using non-standard processing steps. On-wafer inspection is inexpensive and reduces the size of the array system. This shorter optical path length reduces noise from surrounding environments to effectively collect fluorescent signals and reduce the need for conventional optical assemblies for lenses and filters. The quantum efficiency of the photodiode 184 refers to the fraction of photons that are effectively converted into photoelectrons in the active region 185 photons. In terms of standard enthalpy, the quantum efficiency of visible light ranges from 0.3 to 0.5, depending on the root processing parameters such as the amount of cover layer and absorption characteristics. The detection valve of the photodiode 1 84 determines the minimum intensity of the fluorescent light that can be detected. The detection valve 値 also determines the size of the photodiode 1 84 and therefore the number of hybridization chambers 1 in the hybridization and detection section 52 (see Figure 52). The size and quantity of the chambers are limited by the size of the measurement system. The size of the LOC device is the size of the LOC device 301. The size of the LOC device is 301. , 760 micron χ 5,824 micrometers) and technical parameters of the size of available space after intrusion into other functional modules such as pathogen dialysis unit 70 and amplification unit 112. The photodiode 184 detects a minimum of 5 photons in terms of standard 矽 processing. However, to ensure reliable detection, the minimum chirp can be set to 1 光 photons. Thus, in the quantum efficiency range of 0.3 to 0.5 (as discussed above), the fluorescence emission from the probes should be at least 17 photons, but 30 φ photons will contain an appropriate margin of error for reliable detection. Electrochemiluminescence as a Selective Detection Method Electrochemiluminescence (ECL) involves the generation of species on the surface of an electrode that is then subjected to an electron transfer reaction to form an excited state of the emitted light. Electrochemiluminescence differs from normal chemiluminescence in that the formation of the excited species depends on the oxidation or reduction of the luminophore or co-reactant at the electrode. The co-reactant in this case is added to the additional reagent in the ECL solution, which increases the efficiency of # ECL emission. In normal chemiluminescence, the excited species is formed purely by mixing with appropriate reagents. The emitting atom or complex is known as a luminescent group. In short, ECL relies on generating an excitation state of the luminophore in which photons will be emitted. As with any such method, it is possible to take an alternative route from the excited state to not cause the desired light emission (i.e., quenching). The embodiment of the test module using ECL instead of fluorescent detection does not require excitation of the LED. Electrodes are built into the hybrid chambers to provide electrical pulses for generating ECL and to detect such photons using the light sensor 44. The period of the electrical pulse and the voltage are controlled; in some implementations, the control current is selected to control the voltage using -279-201211533. Luminescent Groups and Quenchers The ruthenium complex [Ru(bpy)3]2 + previously described as a fluorescent reporter for such probes can also be used as a luminophore for ECL reactions in the hybridization chamber. TPrA (tri-n-propylamine (CH3CH2-CH2) 3N) was used as a co-reactant. The benefit of the co-reactant ECL is that the luminophores are not consumed after photon emission and the reagents can be reused. In addition, the [Ru(bpy)3]2 + /TPrA E C L system provides good signal levels in aqueous solutions at approximately physiological p Η conditions. Alternative co-reactants, N-butyldiethanolamine and 2-(dibutylamino)ethanol, which produce equal or better results than TPrA and ruthenium complexes. Figure 95 illustrates the reaction that occurs during the ECL process in which [Ru(bpy)3]2+ is a luminophore 864 and the TPrA is a co-reactant 866. The ECL emission 8 62 in the [Ru(bpy)3]2 + /TPrA ECL system occurs after the oxidation of Ru(bpy)32+ and TPrA at the anode 860. These reactions are as follows:

Ru(bpy)32+ -e' ->Ru(bpy)33+ (1) TPrA-e' ^[TPrA*]+ TPrA* + H+ (2)

Ru(bpy)33+ +TPrA* -> Ru(bpy)3*2+ + products (3)

Ru(bpy)3*2+ -> Ru(bpy)32+ +hu (4)

The wavelength of the emitted light 8 62 is about 620 nm' and the anode potential on the Ag/AgCl reference electrode is about 1.1 V. The black hole quencher BHQ RQ previously described as the [Ru(bpy)3]2 + /TPrA -280- 201211533 ECL system will be a suitable quencher. The agent described herein is initially attached to the functional portion of the probe, but in the sample, the quencher may be separated from the solution: hybridization probes for ECL detection are shown in Figures 129 and 130. Hybrid-reactive ECL probes are often referred to as molecular beacons by a single-stranded nucleic acid needle that luminesces when hybridized to a complementary nucleic acid. Figure 129 shows a single ECL probe 237 prior to hybridization of sequence 238. The probe, stem 242, the luminophore 864 at the 5' end, and the loop 240 at the 3' end are bonded together by complementary sequences flanking the sequence of the sequence of the complementary nucleic acid sequence 238 to form a stem complementarity. When the target sequence is absent, the probe is [closed. The stem 242 maintains the luminophore-quenching agent pair. • A significant resonance energy transfer between them can occur, and the ability of the photocell to emit light after electrochemical excitation. Figure 130 shows the ability of the luminescent group 864 and the quencher 248 to spatially separate when the ECL in the open or hybridized configuration hybridizes to the complementary target nucleic acid sequence 23 8 and the light 864 emits light. The ECL emission 862 is used as an indicator that the probe has hybridized. The probe's stem helix hybridized with a highly specific specificity to a complementary target is designed to be quenched in other embodiments of the molecule compared to a non-complementary mono 2 or Iowa black state. Needle 2 3 7. These produced stem loops have a ring 240 quencher 2 4 8 with a target nucleic acid needle. Column composition. The dimensions shown by the probes 242 〇 129 are close to each other to qualitatively eliminate the probe 237. When the structure is destroyed, the hair is recovered optically, because the probe-nuclear probe-281 - 201211533 needle-target helix is more stable. Since the double-stranded DNA is quite robust, the probe-target helix and the stem helix cannot coexist in the stereoscopic space. The stem-loop probe linked to the primer and the linear probe connected to the primer (also known as the scorpion probe) can be used in the LOC device for immediate and Quantitative nucleic acid amplification. The immediate amplification is performed directly in the hybridization chamber of the LOC device. An advantage of using a probe coupled to a primer is that the probe element is actually linked to the primer, so that only a single hybridization event is required during nucleic acid amplification without the need for separate primer hybridization and probe hybridization. This ensures an immediate and efficient response, resulting in a stronger signal, shorter reaction times and better recognition than when using separate primers and probes. The probes (and polymerase and amplification mixture) will be deposited in the hybrid chamber 180 during manufacture without the need to provide an amplification portion on the LOC device. Alternatively, the amplification portion is not used or used for other reactions. Linear ECL probes linked to primers Figures 13 1 and 1 32 show linear ECL probes 693 linked to primers during the first round of nucleic acid amplification, respectively, and primers linked to hybridization during subsequent nucleic acid amplification. Linear probe. Referring to Figure 133, the linear ECL probe 693 coupled to the primer has a double-stranded stem segment 242. One of the probes 696, which is homologous to the region 696 on the target nucleic acid, is linked to the primer, and is labeled with a luminescent group 864 at the 5' end, and is amplified by the 3' end. The fragment 694 is linked to the oligonucleotide primer 700. The stem 242 is -282-201211533 and the other 3' end is labeled with a quencher molecule 248. After completion of the first round of nucleic acid amplification, the probe can be tucked up and hybridized to an extended strand having sequence 69 8 (now complementary). During the first round of nucleic acid amplification, the oligonucleotide primer 700 is affixed to the target DNA 23 8 (see Figure 131) and then extended to form a DNA strand containing both the probe sequence and the amplification product. The amplification blocker 694 prevents the polymerase from reading and replicating the probe region 696. Upon subsequent denaturation, the extended oligonucleotide primer 700/template hybridization system is separated, and the double-strand 242 of the linear probe linked to the primer is also separated, thereby releasing the quencher 248. When the temperature is lowered for the bonding and extension step, the primer-ligated linear ECL probe and the primer-ligated probe sequence 696 are rolled up and hybridized with the amplified complementary sequence 69 8 on the extended strand, and can be detected. Light emission showing the presence of the target DNA is displayed. Linear ECL probes that are not extended and connected to the primer retain their double stems and the light emission remains quenched. This test method is especially suitable for fast detection systems because it requires only a single molecule. Stem-loop ECL probe attached to the primer Figure 133A to 133F show the operation of the stem-loop ECL probe 705 attached to the primer. Referring to Figure 133A, the stem-loop ECL probe 705 attached to the primer has a stem 242 of complementary double stranded DNA and a loop 240 with a probe sequence. The 5' end of one of the stems 708 is labeled with a luminophore 804. Another 710 is labeled with a 3'-end quencher 248 and carries both amplification blocker 694 and oligonucleotide primer 700. During the initial denaturing phase (see Figure 133B), the strands of the target nucleic acid 23 8 are separated, and the stem 242 of the stem-loop ECL probe-283-201211533 705 linked to the primer is also separated. When the temperature is cooled to effect the binding phase (see Figure 133C), the oligonucleotide primer 700 on the stem-loop ECL probe 705 attached to the primer hybridizes to the target nucleic acid sequence 23 8 . During extension (see Figure 133D), the complementary sequence 706 of the target nucleic acid sequence 23 is synthesized to form a DNA strand containing both the probe sequence 705 and the amplification product. The amplification blocker 694 prevents the polymerase from reading and replicating the probe region 705. When the probe is ligated after denaturation (see Figure 133E), the probe sequence (see Figure 133 F) of the loop segment 240 of the stem-loop probe attached to the primer is affixed to the complementary sequence 706 on the extension strand. This configuration is such that the luminophore 864 is at a great distance from the quencher 248, resulting in significantly enhanced light emission. ECL Control Probes Hybridization chamber arrays 110 include some hybridization chambers 180 with positive and negative ECL control probes for analytical quality control. Figures 134 and 135 schematically illustrate a negative control ECL probe 786 that does not contain a luminophore, and Figures 136 and 137 show a positive control E C L probe 7 8 7 without a quencher. The positive and negative control ECL probes have a stem-loop structure as described above for the ECL probe. However, regardless of whether the probes hybridize to an open configuration or remain closed, the positive control ECL probe 787 will emit an ECL signal 862 (see Figure 130), and the negative control ECL probe 78 6 will never emit an ECL. Signal 862. Referring to Figures 134 and I35, the negative control ECL probe 786 does not have a luminescent group (which may or may not have a quencher 248). Thus, whether the target nucleic acid sequence 2 3 8 is hybridized to the probe as shown in Figure 135, or the probe maintains the stem 242 and loop 24 〇 configurations as shown in Figure 34, the ECL signal is 201211533 Ignore. Alternatively, a negative control ECL probe can be designed to remain quenched forever. For example, by having an artificial probe (loop) sequence 240 that does not hybridize to any of the nucleic acid sequences in the study sample, the stem 242 of the probe molecule will self-hybridize itself to render the luminophore and quencher Stay close enough to detect identifiable ECL signals. If the quenching is not complete, this negative control will result in any low volume emissions that may occur. Conversely, the positive control EC L probe 7 8 7 without quencher was constructed as shown in Figures φ 136 and 137. Regardless of whether the positive control probe 787 hybridizes to the target nucleic acid sequence 23 8 , no material quenches the ECL emission 862 from the luminophore 864. Figures 123 and 124 show another possibility to construct a positive control chamber. In this case, the calibration chamber 382, which is closed to the amplicon (or any stream containing the target molecule), can be filled with the ECL luminophore solution so that a positive signal will be detected at the electrode. Likewise, the control chambers can be negative control chambers because of the lack of an inlet® to prevent any target from reaching the probe so that the ECL signal is never detected. Figure 52 shows the positive and negative control probes in the hybridization chamber array 110 ( Possible distributions of 378 and 38 0) respectively. In the case of ECL, the positive and negative control ECL probes 78 6 and 7 87 will be substituted for the control fluorescent probes 3 78 and 380, respectively, and the control probes are placed across the diagonal of the hybrid chamber array 11 In the hybridization room 180. However, the configuration of the control probes within the array is arbitrary (like the configuration of the hybrid chamber array 1 1 )). -285- 201211533 Calibration Chamber for ECL Detection The heterogeneity of the electrical characteristics of the photodiode 184, the reaction to ambient light in the sensor array, and the light originating from the position in the array will be background Noise and offset import output signals. The background is that the calibration chamber in the hybridization chamber array 110 is removed from each output signal, etc. The calibration chamber does not contain any probes, contains no ECL luminophores, or contains illumination that is configured to cause quenching to occur forever. Probes for group and agent. The number of calibration chambers 382 throughout the array of hybridization chambers is arbitrary. However, if the photodiode 184 is calibrated by a relatively close calibration 382, the calibration will be more accurate. Referring to Figure 139, the hybrid column 110 has a calibration chamber 382 for every eight hybridization chambers 180. That is to say, the calibration chamber 382 is placed in the middle of each of the three-by-three hybrid chambers. In this configuration, the hybrid chambers 180 are calibrated by the immediate calibration. Figure 93 shows a differential imager circuit 788 that subtracts the signal differential imager circuit generated from the ECL signal of the surrounding hybridization chamber 180 from the photodiode 184 corresponding to the calibration chamber due to the applied electrical pulse. 78 8 from pixel 790 and "virtual" pixel 792 signal. The signal generated by ambient light in the area of the chamber array is also removed. The signal from pixel 790 is very weak (i.e., close to the dark signal), and it is difficult to distinguish between the background and the very weak number with the reference of the dark signal level. During use, the "read_column" 794 and "read_column_(1" are activated, and the M4 797 and MD4 8 0 1 transistors are turned on. What else is left, the probe is quenched The quasi-room arsenal is also used for the block 382. The sample is taken in the no message 795 switch -286 - 201211533 807 and 809 are turned off so that the output from the pixel 790 and the "virtual" pixel 792 is Stored in the pixel capacitor 803 and the virtual pixel capacitor 850 respectively. After the pixel signal is stored, the switches 807 and 809 are disabled. Then the "read_row" switch 811 and the virtual "read_row" switch 813 are Turn off, the switched capacitor amplifier 815 at the output amplifies the differential signal 8 17 ° φ ECL amount and the signal efficiency ECL efficiency of the normal measurement standard is that each "Faradaic" electron is involved in each electron in the electron The number of photons obtained. The ECL efficiency is expressed by <()ecl:

® where I is the intensity of photons per second, i is the current amperage, F is the Faraday constant, and Να is the Avogadro's constant. The efficiency of the co-reactant ECL is easily eliminated in a deoxygenated, aprotic solution (eg, a nitrogen-laden acetonitrile solution) to allow for efficiency measurements, φΕ (: ι_ consensus 値 approximately 5%. However, direct measurement of the co-reactant The efficiency of the system has been recognized as meaningless. Conversely the 'emission intensity' is related to the results of standard solutions such as Ru(bpy)32+, which are readily prepared in this same form. Literature (see for example j.-287- 201211533 K. Lei and and MJ Powell, J. Electrochem. Soc·, 137, 3127 (1990) and R. Pyati and Μ. M· Richter, Annu. Rep. Prog·Chem.C, 1 03, 1 2-78 (2007)) shows (without enhancers such as surfactants) that the Ru(bpy)32 + ECL efficiency of the TPrA-containing co-reactant is highest (e.g., 2% efficiency) in an amount equivalent to 5% of the ECL elimination in acetonitrile. : See I. Rubinstein & AJ Bard, J. Am. Chem. Soc., 1 03 5 1 2 - 5 1 6 (1 9 8 1 )). ECL potential

The voltage of the working electrode of the Ru(bpy)32 + /TPrA system is about +1,1 V (usually measured in the literature on the reference Ag/AgCl electrode). Such a high electrical compression short electrode life is not a problem for a single use device such as the LO C device used in the diagnostic system of the present invention. The ideal voltage between the anode and the cathode depends on the combination of solution composition and electrode material. Choosing the correct voltage may require compromising between the highest semaphore, reagent and electrode stability, and activation of unwanted side effects such as electrolysis of the water in the chamber. In tests conducted with buffered aqueous Ru(bpy)32+/co-reactant solutions and platinum electrodes, the ECL emission was maximized at 2.1-2.2 V (depending on the co-reactant selection). When the voltage is lower than 1.9 V and higher than 2.6 V, the emission intensity drops to a maximum of < 75%. When the voltage is lower than 1.7 V and higher than 2.8 V, the emission intensity drops to a maximum of < 50%. Therefore, the preferred anode-cathode voltage difference for such ECL operating systems is from 1.7 to 20.8 V, particularly preferably in the range of 1.9 to 2.6 V. This allows to maximize the emission intensity as a function of voltage while avoiding the observation of significant gas evolution at the electrodes. -288- 201211533 ECL Emission Wavelength The wavelength of the emitted light 862 from the ECL is at a peak of about 620 nm (measured in air or vacuum) that spans a fairly broad wave. Significant emissions occur at wavelengths between about 550 nm and 700 nm. The peak emission wavelength can be varied by about 10% due to chemical cycling around the active species. The LOC device described here does not have a specific wavelength filter, so that such a broad and varied spectrum has two advantages. This first advantage is sensitivity: any filter reduces light penetration, even within the passband, so efficiency can be improved by the filter. The second advantage is adaptability: there is no need to adjust the filter passband after a small number of trials, and the signal is less dependent on the slight difference in the non-standard components of the input. Solution Volume Participating in ECL The availability of luminophores (and co-reactants) in the ECL-dependent solution ® ' As shown in Figure 97, the excited species 868 is only produced in solution 872 near the electrode 87 。. The depth of the reference layer in the model shown here is the depth of the layer 872 of the solution around the electrode 860, and the excited species 868 is produced. This is a simplified diagram because solution kinetics can drive the available concentrations down: • Increase usability: diffusion and electrophoresis effects will allow for more exchanges • Reduced usability: reagents can be adsorbed on the electrodes and become strong and long range The change of the implementation state captures the signal length including the change of the agent into the sample. However, the 860 and the number of boundaries where the above or more solutions cannot be used by the -289-201211533 ECL process. With the boundary layer depth of 値〇·5 μm, the following observations were made: ECL was observed in an experiment in which a magnetic bead conjugate with a diameter of up to 4.5 μm was used to attract the luminophore 864 to the anode 860.

The Ru(bpy)32 + /TPrA ECL emission 862 varies with electrode spacing and is found to be maximized at 0. 8 micron electrode spacing for the interdigitated electrode array. When the electrode spacing is about 2 microns, the need for a co-reactant 866 in the aqueous solution 826 can be removed. This shows that the excited species 8 68 diffuses a few microns, suggesting that the species in the ground state are diffusion exchanged on a similar scale. Steady State and Pulse Operation During pulse activation of the electrodes 860 and 870, the intensity of the ECL emission 862 (see Figure 130) is generally higher than the intensity of the emission 8 62 from the steady state activation of the electrodes. Thus, the activation signals to the electrodes 860 and 870 are pulse width modulated (PWM) by the CMOS circuit 86 (see Figure 102). 〇Reagent recycling and species lifetime The Ru complex will not be in RU(bpy) The 32 + /TPrA ECL system is consumed, so the intensity of this emission 862 does not decrease with the continuous reaction cycle. The life of the rate limiting step is about 0. 2 milliseconds, so the total reaction cycle time is about 1 millisecond. Electrophoretic effects and other limitations -290- 201211533 Due to the complexity of the solution in the hybrid chamber, many phenomena occur when the ECL voltage is turned on. Electrophoresis of large molecules, ohmic conduction, and capacitive effects from small ion movements occur simultaneously. Electrophoresis of oligonucleotides (probes and amplicons) complicates the detection of the probe-target hybrid because the DNA is highly negatively charged and attracted to the anode 860. The time for this action is usually very short (in milliseconds). Even if the voltage is only moderate (1 V), the electrophoretic effect is still strong because the interval between the anode 860 and the cathode φ 87 很小 is small. Electrophoresis increases the ECL emission 826 in some embodiments of the LOC device, which is reduced in other embodiments. This is achieved by increasing or decreasing the electrode spacing to achieve this correlation increase or decrease the electrophoretic effect. The anode 8 60 and cathode 870 above the photodiode 184 represent the extremes of minimizing this spacing. This arrangement produces an ECL even without co-reactants 8 66 at carbon electrodes 8 60 and 870. # 欧姆加热 (DC Current) The current required to maintain the ECL voltage of approximately 2.2 V is determined as follows, with reference to ECL cells 874 as illustrated schematically in Figure 98. The DC current through the chamber is determined by two resistances: the interfacial resistance Ri between the electrodes 8 60 and 8 70 and the body of the solution, and the solution resistance Rs derived from the bulk solution resistivity and conduction path geometry. In the case of a solution having an ionic strength associated with the conditions in the LOC device, the chamber resistance is dominated by the interface resistance at the electrodes 860 and 870, and Rs can be ignored -291 - 201211533 The effect of the interface resistance is passed through The calibration of the macroscopic current of a similar solution of the electrode geometry in the LOC device is estimated. A giant measurement of the current density through a similar solution at the platinum electrode is obtained. Consistent with the worst case (high current) employed, the overall ionic strength and ECL reactant concentration in the test solution is higher than that of the user in the LOC device. The anode region is smaller than the cathode region and is surrounded by a cathode having a relatively large area of annular geometry. In the case of an anode consisting of a circle having a diameter of 2 mm, the measured current is 1.1 m A, resulting in a current intensity of 350 A/m2. In the heating model, the electrode zone is illustrated as a square annular geometry as illustrated in Figure 98. The anode is a ring that is 1 micron wide and 1 micron thick. The surface area is 196 square microns, so the calculated current is 1 = 69 nA. Heating (power = V2/R) is modeled on the worst case where all of the heat is used to increase the temperature of the water in the chamber. This results in heating the contents of the chamber at 5.8 °C / s with a voltage difference of 2.2 V, if the heat removed by the LOC device body is not corrected. Heating the chamber at about 20 °C can cause most of the hybrid probe to denature. For highly specific probes for mutation detection, it is preferred to further limit the heating to 41 or less. With this degree of temperature stability, it is possible to make a single base mismatched sensitive hybridization with a suitably designed sequence. This allows for the detection of mutations and allelic differences at the level of single nucleotide polymorphism. The DC current was therefore applied to electrodes 860 and 8 70 for 0.69 seconds to limit the heating to 4 °C. A current of about 69 nA passes through the chamber far beyond the Faraday current that can be accommodated by the ECL species at a micro-292 - 201211533 molar concentration. Thus, pulsing the electrodes 860 and 870 with a low duty cycle further reduces heating (to i ° C or less) while maintaining sufficient ECL emission 862 without causing side effects associated with reagent consumption. In other embodiments, the current is reduced to 0.1 nA, which removes the need for the electrodes to activate the electrodes. Even with currents as low as 0.1 nA, the ECL emission 8 62 is limited to luminophores. φ chamber and electrode geometry Maximize the optical coupling between the ECL luminescence and the light sensor. The direct chemical pre-system of ECL luminescence is generated within a few nanometers of the working electrode. Referring again to Figure 97, light emission (the excited species 868) typically occurs within a few microns or less of the location. Thus the volume adjacent to the working electrode (anode 8 60) can be seen by the corresponding photodiode 184 of the light sensor 44. In view of this, the electrodes 860 and 870 are in close proximity to the active surface region 185 of the corresponding photodiode 184 in the photosensor 44. In addition, the shape of the anode 8 60 is structured to increase the side periphery thereof as seen by the photodiode 184. The purpose of this is to maximize the volume of the excited species 868 that can be detected by the lower photodiode 184. Figure 96 schematically illustrates three embodiments of the anode 860. The comb structure anode 878 has the advantage that the parallel finger configuration 880 can intersect the finger configuration of the cathode 870. The interdigitated configuration is shown in Figures 1 and 3, and the LOC portion configuration as shown in Figures 120 and 124. The interdigitated configuration provides a relatively narrow (1 to 2 micron) uniform dielectric gap 876 (see Figure 97), and the interdigitated comb structure is relatively simple for the lithography manufacturing process. As discussed above - 293-201211533, a relatively narrow dielectric gap 8 76 between the electrodes 860 and 870 eliminates the need for co-reactants in the solution 872 because the excited species 868 will diffuse between the anode and the cathode. . The need to remove the co-reactant removes the potential chemical effects of the co-reactant on various detection chemistries and provides a wider range of possible detection options. Referring again to Figure 96, some embodiments of the anode 860 have a bent configuration 882. In order to achieve a high perimeter length while maintaining tolerance to manufacturing errors, it is convenient to form a wide, rectangular corner 884. φ The anode can have a more complex configuration 886 if needed or desired. For example, it may have a toothed configuration 880, a branching structure 8 90, or a combination of the two. A partial diagram of the LOC design with branch structure 8 90 is shown in Figures 138 and 139. This more complex configuration, such as 8 86, provides a long-length side perimeter that is best suited for solution chemistry where co-reactants are used, since patterning closely spaced opposite cathodes is more difficult. Electrode thickness ♦ Typically, ECL cells are involved. The working electrode is a flat surface from the outside. In addition, micro-fabrication techniques conventionally used for metal layers tend to result in planar structures having a metal thickness of about 1 micron. As previously described and shown in Figures 96, 99 and 1 ,, increasing the length of the side perimeter promotes coupling between the ECL emission and the photodiode 1 84. A second strategy to further increase the efficiency of collecting the emitted light 862 (see Figure 130) by the photodiode 184 is to increase the thickness of the anode 860. This illustration is shown in Figure 97. The portion of the participating volume 892 adjacent to the wall of the working electrode 8 201211533 is the region where the photodiode 184 is most effectively coupled. Thus, with a given width of working electrode 860, the overall collection efficiency of the emitted light 862 can be increased by increasing the thickness of the electrode. In addition, the width of the working electrode 860 is reduced as much as possible because of the high current required to carry the capacitor. The thickness of the electrodes 860 and 870 cannot be increased without limitation. It is noted that the characteristics and spacing of the electrodes may be on the order of 1 micron, and liquid charging makes the gap larger than the depth unfavorable, so that the ideal thickness of the electrodes is 0.25 to 2 micrometers. Electrode Gap The gap between electrodes 860 and 870 is very important for the signal quality in the LOC device, particularly in the implementation of the electrodes interdigitated. In embodiments in which the anode 8 60-branch structure, such as shown in Figures 96 and 00, the gap between adjacent elements is also very important. Both the ECL emission efficiency and the collection efficiency of the emitted light should be maximized. # Generate ECL emission propensity to prefer an electrode gap of 1 micron or less. Small gaps are particularly attractive when ECL is carried out in the absence of a co-reactant. The fact that the gap can correspond to the wavelength of the emitted light 8 62 is not of importance. Thus, in many embodiments where the emitted light 862 (see Figure 130) is in a positional measurement where the light is not required to pass between the electrodes 860 and 870, making the electrode gap as small as possible is generally a goal. However, in an embodiment in which the emitted light 826 has to pass between the electrodes 860 and 870, it becomes necessary to consider not only the ECL emission process but also the wavelength property of the light. -295- 201211533 The emission of light from the Ru(bpy)32+ ECL 8 62 is about 620 nm, so it is 460 nm (0.46 μm) in water. In the embodiment in which the photodiode 184 and the ECL excitation species 6.8 are located on different sides of the electrode structure, and the electrode structure is a metal, the emitted light 8 62 must pass through the metal structural elements. The gap between them. If the gap is the same as the wavelength of the light, the diffraction generally reduces the intensity of the propagating light reaching the photodiode 184. However, if the emitted light 862 is incident on the gap at a large angle, the subtractive mode coupling can be utilized to enhance the intensity of the collected signal. Two methods are employed in the LOC device to increase the coupling efficiency between the photodiode 184 and the emitted light 826. First, the gap between the metal elements is not reduced to about the wavelength of the emitted light below the water, i.e., about 0.4 microns. When combined with other observations regarding the small gap between the finger electrodes, this shows that the optimum range of the electrode spacing is 0.4 to 2 microns. Second, the distance between the elements to the photodiode 180 is minimized in the LOC device implementation described herein, which is shown at the electrodes 8 60 and 807 and the photodiode The total thickness of the layers between the polar bodies 184 is 1 micron or less. In embodiments in which there are multiple layers between the electrodes and the photodiode, arranging their thickness to a quarter-wavelength or three-quarters wavelength layer has the added benefit of suppressing reflection of the emitted light 862. Electrode Model Figure 97 is a partial cross-sectional view of electrodes 860 and 870 in a hybridization chamber -296-201211533. The volume around the side periphery of the anode 860 is occupied by the excited species 868, which is sometimes referred to as the participating volume 892. The enclosed region 894 above the anode 860 is ignored because its optical coupling with the photodiode 184 can be ignored. Techniques for determining whether a particular electrode configuration is the underlying photodiode 184 providing an ECL emission 862 will be described below with reference to Figures 98, 99 and 100. φ Figure 98 is an annular configuration in which the anode 860 is located around the edge of the photodiode 184. In Fig. 99, the anode 860 is located within the periphery of the photodiode 184. Figure 1A shows a more complex configuration in which the anode 860 has a series of parallel finger-like configurations 800 to increase the length of the sides. For all of the above configurations, the model calculation is as follows. In terms of the participating volume 892 of the solution VECL, the total effective number of the emitter Nem is: ^ Nem = Nium. τρ / Tecl = VeclClNa . Tp/TECL (6) where the number of participants of the luminophore Nlum = VECLCLNA, r ECL is the ECL process Lifetime, CL-based luminophore concentration, rp-pulse period, and Na-type Avogador's number. Number of photons emitted by isotropic emission Nph()t: (7)

Nph〇t = φε〇ί Nem -297- 201211533 wherein φΕα-based ECL efficiency is defined as the average number of photons emitted by the ECL reaction of a single luminophore. Then, the number S of electronic signals from the photodiode is S=Nph〇t. φοφς, (8) where Φ. The optical coupling efficiency (the number of photons absorbed by the photodiode 184) and the quantum efficiency of the photodiode. Therefore the signal is:

TECL is in the electrode configuration of Figures 98 and 99, φ. Department: ()). == (25% of photons directed to photodiode 84) χ (10% of unreflected photons) is the configuration (|) shown in Figures 98 and 99. = 2.5% In the electrode configuration shown in Fig. 100, 50% of the photons are emitted to the direction of 0 to the photodiode 184, but the function of the absorption efficiency for the angle is constant and therefore small. = (50% of the photons directed to the photodiode) χ (10% of the unreflected photons) ie the configuration shown in Figure 100 = 5 % The participating volume 8 92 depends on the electrode configuration, a detailed description In the corresponding chapter. The input parameters used for the calculation are listed in the following table: -298- 8 201211533

Table 5: Input Parameter Number of Parameters 値 Note Luminescence Concentration Cl 2.89 μιη Previously Calculated Probe Concentration ECL Recycling Period (Lifetime) rECL 1 ms Combination Life of Luminescence Reaction Step Boundary Depth D 0.5 μπι Participation in ECL Solution Effective volume (including diffusion and electrophoresis) During application of current rp 0.69 s is selected to limit ohmic heating to 4 ° C (as previously described) Chamber X size 28 μιη Room Y size 28 μπα Chamber height Z 8 μιη Photodiode X Size 16 μπι Photodiode Y size 16 μιη electrode thickness (ie exposed edge height) 1 μηι Electrode layer minimum width and gap 1 μιη Process critical dimension electrode interface current density 350 A/m2 for ohmic heating solution volume resistivity 0.5 Qm is used for ohmic heating applied voltage difference (working-opposite electrode) 2.2 V ring geometry around the photodiode Referring to Figure 98, the anode 860 is a ring around the edge of the photodiode 184. In this configuration, the participating volume 892 is:

Vecl = 4x [(the layer beside the electrode wall) + (a quarter of the cylinder on the electrode wall)] Calculation result: Photons generated from the 0.5 micron boundary layer: 3.lxlO5 electrons in the photodiode Number: 2.3χ103 This signal can be easily detected by the lower photodiode-299-201211533 body 184 of the LOC device light sensor 44. Additional Finger Shape Construction to Increase Edge Length Referring to Figure 100, a parallel finger configuration 880 is applied throughout the anode 860. Only the horizontal edges shown in the figure contribute to the participating volume 892 to avoid double counting the vertical edges. Thus the participating volume 8 92 is: VECL = (8x2) x [(layer next to the electrode wall) + (quarter of the cylinder on the electrode wall)] Figure 100 configuration calculated as: From 0.5 micron boundary Photons generated by the layer: l.lxlO6 Electron number in the photodiode 184: 8.0 X 1 03 This signal can be easily detected in the photodiode 184. The complete coverage shown in Figures 101 and 102 is included as a limit for maximum surface area coupling. In fact, the electrode surface area is 90% between the active surface area 185 of the photodiode 184 or Better coupling achieves nearly optimal results, even though the photodiode active surface region 185 is 50% coupled to the electrode surface region to provide most of the benefits of the fully covered configuration. The complete coverage can be achieved in two implementations: first, as shown in FIG. 101, by using a transparent anode 860 on a plane parallel to the photodiode 184 and matching the area of the transparent anode to the photodiode The area of the pole body is arranged adjacent to the photodiode 1 84 such that the emitter -300 - 201211533 light 8 62 passes through the anode to the photodiode. In the second embodiment shown in FIG. 102, the anode 860 is also parallel and registered with the photodiode region, but the solution 8 72 is filled with the anode 860 and the photodiode 184. Space between. In the case of a signal model of the full coverage configuration, assuming that the anode is a complete layer above the photodiode 184, half of the photons are directed to the photodiode 184 (absorption efficiency is still 10%). Photons generated from a 0.5 micron boundary layer: 7.7xl05 φ Number of electrons in the photodiode: 1.2x1 04 By using a surfactant and a probe fixed to the anode, it is possible to enhance the signal and in addition to the above model. Detection. Maximum separation between the ECL probe and the photodiode The detection of hybridization on the wafer does not require detection with a conjugated focal microscope (see prior art). This difference from traditional inspection techniques represents an important factor in saving time and cost. Traditional inspections require imaging optics that use lenses and curved mirrors. By employing non-imaging optics, the diagnostic system avoids the need for complex and cumbersome strings of optical components. The placement of the photodiode in close proximity to the probe has the advantage of extremely high collection efficiency: when the material thickness between the probe and the photodiode is 1 micron, the emission angle of the emitted light is as high as 174°. . This angle is calculated by considering the light emitted from the probe closest to the center of the surface of the hybridization chamber of the photodiode, the photodiode having a planar active surface parallel to the surface of the chamber. Light within the light emitting pyramid can be absorbed by the photodiode, which is defined as having a firing probe at its apex and at the sensor corners around its plane. -301 - 201211533 For example, a 16 micron χ 16 micron sensor has an apex angle of 170. The apex angle is 174° in the extreme case where the photodiode is expanded such that its area conforms to the 28 micron x 2 6.5 micron hybrid cell area. It is easily achieved that the interval between the surface of the chamber and the active surface of the photodiode is 1 micron or less. The application of non-imaging optical methods does require that the photodiode 184 be placed very close to the hybridization chamber to collect a sufficient amount of fluorescent emission photons. The maximum spacing between the photodiode and the probe is determined as follows. Using the ruthenium chelate luminophore and the electrode configuration of Figure 100, we calculated that 27,000 photons were absorbed from our respective hybridization chambers 180 by our 16 micron X 16 micron sensor to produce 8,000 electrons (assuming sensor quantum The efficiency is 30%). In doing this calculation, we assume that the light collection area of the hybridization chamber has the same bottom area as our sensor, and that a quarter of the total number of hybrid photons is adjusted to reach the sensor and is conservative It is estimated that 1 〇% of the photon ratio will not be scattered from the sensor interface. That is, the light collection efficiency of the optical system is φο = 〇.025. Lu more precisely, we can write <h = [(the bottom area of the light collection area of the hybridization chamber) / (photodetector area)] [Ω / 4 7 Γ ] [10% absorption], where Ω = The solid angle to which the photodetector of the representative point on the substrate of the hybridization chamber opposes. In terms of the geometry of the regular square pyramid: Q = 4arcsin(a2/(4d〇2 + a2)), where d〇 = the distance between the chamber and the photodiode, and a is the size of the photodiode . Each hybridization chamber releases l.lxl 〇6 photons. The selected photodetector has a detection low limit of 17 photons, which is greater than the dG値 of the sensor size of 10×302-201211533 (ie, substantially perpendicular incidence), and the non-reflection is in the The proportion of photons in the face can be increased from 10% to 90%. Therefore, the required optical efficiency is: φ 〇 = 17 / (1.1 χ 106 χΟ. 9) = 1.72 x 10 '5 The light-emitting region of the hybridization chamber 180 has a bottom area of 29 φ 19.75 μm. Decomposing dQ will result in a distance dQ = 1,600 microns between the hybridization chamber and the photodetector. Under this limitation, the taper angle as defined above is only 〇. 8°. It should be noted that this analysis ignores the negligible negligible LOC variant. All of the LOC devices 30 1 described and illustrated above are only one of the LOC device designs. Variations from the sample inlet to the detection using the LOC devices of the various functional combinations described above will now be added and/or shown in a schematic flow diagram to demonstrate some possible combinations of flow charts that are divided into sample input and preparation stages as appropriate. 290. Culture stage 291, amplification stage 292, and pre-hybridization stage detection stage 294. For all such LOC variants that are only briefly described or indicated, their complete configuration is not shown to be concise. Also for the sake of clarity, smaller functional units such as liquid and temperature sensors are not shown, but it should be understood that the impact of the minimum micron X maximum limit of these single detector tables may vary. Description. These extraction steps 293 and the intention to show that the somatosensory position has been broken into the appropriate positions in the following LOC device designs by -303- 201211533. LOC Device with ECL Detection Figures 111 through 127 show LOC Variant 729 with Electrochemiluminescence (ECL) detection. The LOC device prepares 288, extracts 290, cultures 291, expands 292, and detects nucleic acids of 294 humans and pathogens, as well as protein detection of humans and pathogens. ECL is used in hybrid chamber arrays and protein body assay chamber arrays to detect targets. Preferably, as shown in Figure 117, a biological sample (e.g., whole blood) is added to the sample inlet 68. The sample flows through the capping channel 94 to the anticoagulant surface tension valve 118. The upper cover 46 is constructed to have an interface layer 594 (see FIG. 112) between the upper cover channel layer 80 and the MST channel layer 100 of the CMOS + MST device 48. The interfacial layer 594 allows for more complex fluid intercommunication between the reagent reservoirs and the MST layer 87 without increasing the size of the crucible substrate 84. Figure 113 shows the MST layer 87 that can be seen on the upper surface of the CMOS+MST device 48. Figure 114 shows the cover channel layer 80 on the underside of the upper cover 46. Figure 115 overlays the reservoirs, the upper cover channels 94 and the interface channels to illustrate the more achieved by the upper cover 46 of the interface layer 594. For complex channel systems. Preferably, as shown in FIG. 117, the interface layer 594 requires the anticoagulant surface tension valve 118 to have two interface channels 596 and 598. The reservoir side interface channel 5 96 is connected to the outlet of the reservoir and the descending port 92 and the sample side interface channel 598 are connected to the rising port 96 and the upper cover channel 94° -304 - 201211533. The anticoagulant from the reservoir 54 is passed through the reservoir. The device side interface channel 596 flows through the MST channel 90 to form a meniscus at the riser 96. The sample stream in the capping channel 94 is lowered to the sample side interface channel 598 to remove the meniscus to combine the anticoagulant with the blood sample on the way to the leukocyte dialysis section 328. The white blood cell dialysis section 328 has a bypass channel 600 for accommodating the flow channel structure without trapping air bubbles (see Figures 117 and 126). The blood sample φ flows through the upper cover passage 94 to the upstream end of the large component interface passage 730. The large component interface channel 730 is in fluid communication with the dialysis MST channel 204 via a hole in the form of a 7.5 micron diameter hole 165 (see Figure 126). Referring to Figure 126, each of the dialysis MST channels 206 is from the 7.5 micron diameter hole 165 to a respective dialysis riser 168. The dialysis rise ports 1 68 are open to the small component interface channel 73 2 . However, the risers are configured to hold the meniscus rather than allowing the capillary drive flow to continue. The riser port of the bypass channel 600 has a capillary initiation feature 202 that is configured to initiate a capillary drive flow into the small component interface channel 732. This ensures that the flow begins at the upstream end of the small component interface channel 732 and sequentially releases the meniscus at the dialysis riser 168 as the flow progresses downstream. Figure 121 shows the downstream end of the white blood cell dialysis section 328. The large component interface channel 73 0 feeds into the large component upper cover channel 736, and the small component interface channel 732 feeds into the small component upper cover channel 734 » as best shown in FIG. 115, the large component upper cover channel 736 feeds white blood cells (and any other large components) to the chemical lysis portion 130.1 via the lysis surface tension valve 128.1, and dissolves from the reservoir 56.1 - 305 - 201211533 at the lysis surface tension valve 128. Cell reagent. The chemical lysis section 130.1 has a 3 micron filtration drop port 7 3 8 at the outlet (see Figure 17). The filter lowering port ensures that no large component reaches the boiling start valve 206 of the lysate outlet. After a sufficient time, the boiling start valve 206 opens the outlet of the chemical lysis unit 1 30. 1 , the sample flow is divided into two paths. flow. Preferably, as shown in Figure 117, all the way to the surface tension valve 132.1 of the first restriction enzyme, ligase and linker reservoir 58.1, and the other route is directly attracted to the hybrid along the lytic leukocyte channel 742. And the protein body detection chamber array 1 24.1 in the detecting unit 294. Here the sample is filled with the protein body detection chamber array 124.1 (see Figure 19), which contains a probe for hybridization with the target human protein. The probe-target hybridization system is detected by light sensor 44 (see Figure 1 1 1). This other route flows into the white blood cell culture section 114.1 together with the restriction enzyme, ligase, and linker primer from the reservoir 58.1. Referring to Figure 181, after restriction enzyme digestion and linker ligation, the culture outlet valve 207 (also a boiling start valve) is opened and fluid continues to advance to the white blood cell DNA amplification portion 112.1. The amplification mixture and polymerase in reservoirs 60.1 and 62.1 were added via surface tension valves 138.1 and 140.1, respectively. Referring to Figure 119, after the heating cycle, the boiling activation valve 108 is opened to allow the amplicon to enter the hybridization chamber array 110.1 containing probes for human DNA targets. The probe-target hybridization system is detected by the light sensor 44. Red blood cells and pathogens from the leukocyte dialysis section 3 28 are fed to the pathogen dialysis section 70 via the capping channel 73 4 (see Figs. 117 and 127). This operates in the same manner as the white blood cell dialysis section 328 except that the -306-201211533 filter down port has a 3 micron aperture 164 rather than a 7.5 micron aperture 165 used for leukocyte dialysis. The red blood cells are maintained in the large component interface channel 730 and the pathogen diffuses to the small component interface channel 73 2 . Figure 122 shows the downstream end of the pathogen dialysis section 70. The red blood cells flow into the large component capping channel 736 which fills the small component capping channel 734. It will be understood that "large components" and "small components" are used in a relative sense because the large component output of the pathogen dialysis section is part of the small component output of the white blood cell φ dialysis section. The components of the large component upper cover 73 6 or interface channel within the particular dialysis section are simply larger than the components of the small component upper cover 734 or interface channel. Preferably, as shown in Figures 1 1 5 and 16 16, the red blood cells in the large component upper cover channel 73 6 are directed to the surface tension valve 128.3 of the lysis reagent reservoir 56.3. The lysis reagent is combined with the red blood cells when the sample fluid is filled with the chemical lysis portion 130.3. The boiling start valve 206 at the exit of the third chemical lysis section 130.3 retains the pathogen until the lysis is complete. When the boiling start valve 206 is opened, the red blood cell DNA # flows directly into the protein body detection chamber array 124.3 for protein analysis and detection by the light sensor 44 (see Fig. 19). The pathogen in the small component capping channel 734 is directed to the surface tension valve 128.2 of the second lysis reagent reservoir 56.2. The lysis reagent is combined with the pathogen when the sample fluid is flooded with the second chemical lysis portion 130.2. After sufficient time, the boiling start valve 206 opens the outlet of the chemical lysis section 130.2, and the sample stream is split into two paths. Preferably, as shown in Figures 116 and 118, one way flows to the surface tension valve 132.2 of the second restriction enzyme, ligase and linker 58.2, and the other path is directly attracted to the side channel 744 by -307-201211533. Hybridization and detection unit 294. Here the sample is filled with the protein plastid detection chamber array 124.2 (see Figure 119) containing probes for hybridization with the target pathogen protein or other biomolecule. The probe-target hybridization system is detected by the light sensor 44 (see Figure 1 1 1). This other route flows into the pathogen culture unit 1 1 4.2 together with the restriction enzyme, ligase and linker primer from the reservoir 58.2. After restriction enzyme digestion and linker ligation, the culture outlet valve 207 (also a boiling start valve) is opened and fluid continues to advance to the pathogen DNA amplification portion 112.2 (see Figure 118). When the chamber is filled, the amplification mixture and polymerase in reservoirs 60.2 and 62.2 are added via surface tension valves 138.2 and 140.2, respectively. After the heating cycle, the boiling start valve 108 is opened to allow the amplicon to flow into the second hybridization chamber array 1 10.2 containing the probe for the pathogen DNA target. The probe-target hybridization system is detected by the light sensor 44 (see Figure 119, referring to Figure 120, the hybridization chamber arrays 110.1 and 110.2 and the protein body detection chamber arrays 124.1 to 124.3 are made of strips of titanium nitride. The heater element 1 8 2 » is detected by the end liquid sensor 178 when the fluid reaches the end of the hybrid chamber array or the protein body detection chamber array, and the heater 108 is then activated after a delay. A flow rate sensor 74A (see Figure 125) is included in the pathogen culture portion 114.2 to determine the time delay. Figures 123 and 124 show the calibration chambers 382. They are used to calibrate the photodiode 1 84 to adjust System noise and background 値. The response and electrical noise characteristics of the photodiode can vary with position and heating. -308- 201211533 The output signal from the calibration chamber 382 without any probe is closely related to Noise and background in the output signals of all chambers. Subtracting the calibration signal from other hybrid chambers removes the noise substantially to remove the noise, leaving the signal generated by the chemiluminescence (if any). Ground The positive and negative control ECL probes 786 and 787 can be placed in some of the hybridization chambers 180 for analytical quality control. Referring to Figure 116, the phisizer 196 consisting of a reservoir 188 and an evaporator 190 is located at the upper left of the device. The humidity sensor 232 is located adjacent to the hybridization chamber array 1 1 , where the humidity measurement is important to reduce the evaporation of the solution containing the exposed probe. The dialysis portion is output by combining the white blood cells and the pathogen to generate three Outflows (white blood cells, red blood cells and pathogens and other biomolecules), the three output streams are treated separately to allow for higher sensitivity and parallel analysis. The output of each stream is dissolved and separately directed to the protein body detection chamber The array is used for detecting proteins. The dissolved white blood cells and pathogens are also directed to the culture portion 1 14 and the amplification portion 1 1 2 for amplification 'and then hybridized for nucleic acid detection. The hot groove LOC device is preferably as shown in Fig. 1 2 8 , and the groove 869 is etched to the back of the ruthenium substrate. The purpose of the groove is to thermally isolate the amplification portion 1 1 2 from the hybridization chamber column 1 1 0. The hybrid array Contains a detection probe that is degradable at high temperatures. The trench has a thermal conductivity less than 6,000 times that of the tantalum substrate when filling the air' by significantly reducing the heat flux entering the adjacent portion of the LOC device. The input of the white column is made up of 84 arrays. This - 309-201211533 provides two main advantages: increasing the heating efficiency in the amplification section 1 1 2; and reducing the undesired temperature rise in the adjacent hybridization section 1 1 〇 Improving the heating efficiency means that less power is required to heat the amplifying portion 112 and the temperature reaches the desired end temperature more quickly and more spatially consistent within the amplifying portion. Reducing the temperature rise in the hybridization section 110 allows for a wider range of probe chemistry and superior signal quality. The trench can be placed around any area on the LOC device to thermally isolate components in the region. The width and depth of the groove 896 can be varied to suit the particular application. Conclusion The devices, systems, and methods described herein facilitate rapid, inexpensive, and suitable molecular diagnostic testing for local care. The above-described system and its components are merely illustrative, and many variations and modifications will be readily apparent to those skilled in the art without departing from the scope of the invention. BRIEF DESCRIPTION OF THE DRAWINGS The preferred embodiments of the present invention will now be described by way of example only with reference to the accompanying drawings in which: FIG. 1 shows a test module and test module configured for fluorescence detection. Figure 2 is a schematic diagram of the electronic components in the test module configured for fluorescence detection; Figure 3 is a schematic diagram of the electronic components in the test module reader; -310- 201211533 Figure 4 is a schematic view of the structure of the LOC device; Figure 5 is a perspective view of the LOC device; Figure 6 is a plan view of the LOC device, the features and structures of all layers are overlapped with each other; Figure 7 is a plan view of the LOC device, which is separate Figure 8 is a top view of the upper cover, with internal lines and receptacles shown in dashed lines; • Figure 9 is an exploded top view of the upper cover, with internal lines and receptacles shown in dashed lines: Figure 10 is a lower view of the upper cover, Display the configuration of the upper channel;

Figure 11 is a plan view of the LOC device, showing the structure of the CMOS + MST device independently; Figure 12 is a schematic cross-sectional view of the sample inlet of the LOC device; Figure 13 is an enlarged view of the AA region shown in Figure 6; Figure 14 is Figure 6 Figure 15 is an enlarged view of the AE area shown in Figure 13; a partial perspective view of Figure 16 illustrates the layered structure of the LOC unit in the AE area; Figure 17 is a partial perspective view illustrating the AE The layered structure of the LOC device in the zone; the partial perspective view of Figure 18 illustrates the layered structure of the LOC device in the AE zone; the partial perspective view of Figure 19 illustrates the layered structure of the LOC device in the AE zone; -311 - 201211533 A partial perspective view of Figure 20 illustrates the layered structure of the LOC device in the AE area. A partial perspective view of Figure 21 illustrates the layered structure of the LOC device in the AE area: Figure 2 2 is a diagram of Figure 2 A schematic cross-sectional view of the lysis reagent reservoir; a partial perspective view of Figure 23 illustrates the layered structure of the LOC device in zone AB; and a partial perspective view of Figure 24 illustrates the layered structure of the LOC device in zone AB: Figure 25 The perspective view illustrates the layered frustration of the LOC device in the AI zone. The partial perspective view of Figure 26 illustrates the LOC device in the AB zone. Layered structure; Figure 27 is a partial perspective view showing the layered structure of the LOC device in the AB zone; Figure 28 is a partial perspective view showing the layered structure of the LOC device in the AB zone; Figure 29 is a partial perspective view illustrating the AB zone Figure 30 is a schematic cross-sectional view of the augmentation mixing reservoir and the polymerase reservoir; Figure 31 shows the characteristics of the boiling start valve independently; Figure 32 is a schematic cross-sectional view of the boiling start valve, the cross-section along the figure Figure 33 is an enlarged view of the AF area shown in Figure 15; -312-201211533 Figure 34 is a schematic cross-sectional view of the upstream end of the dialysis section, the section along Figure 3 Figure 35 is an enlarged view of the AC region shown in Figure 6; Figure 36 is a re-enlarged view of the AC region, which shows the amplification portion; Figure 37 is the AC region again. Enlarged view, the figure shows an amplifying part; Fig. 38 is a re-enlarged view of the AC area, which shows an amplifying part; Figure 39 is a re-enlarged view of the AK area shown in Figure 38;

Figure 40 is a re-enlarged view of the AC region, which shows an amplification chamber; Figure 41 is a re-enlarged view of the AC region, which shows an amplification portion; Figure 42 is a re-enlarged view of the AC region, which shows an amplification chamber Figure 43 is a re-enlarged view of the AL region shown in Figure 42; Figure 44 is a re-enlarged view of the AC region, which shows an amplification portion; Figure 45 is a re-enlarged view of the AM region shown in Figure 44; A re-enlarged view of the AC region, which shows an amplification chamber; Figure 47 is a re-enlarged view of the AN region shown in Figure 46; Figure 48 is a re-enlarged view of the AC region, which shows an amplification chamber; A re-enlarged view of the AC region, which shows an amplification chamber; Figure 50 is a re-enlarged view of the AC region, which shows an amplification portion; Figure 51 is a schematic cross-sectional view of the amplification portion; Figure 52 is an enlarged plan view of the hybridization portion; Figure 53 is a re-enlarged plan view of two independent hybridization chambers; Figure 54 is a schematic cross-sectional view of a single hybridization chamber; Figure 55 is an enlarged view of the humidifier in the AG region shown in Figure 6; Figure 56 is a diagram of Figure 52 Enlarged view of the AD area; -313- 201211533 Figure 57 is an exploded perspective view of the LOC device of the AD area; Figure 58 is an enlarged plan view of the humidity sensor shown in the AH area of Figure 6, Figure 59 A cross-sectional view of the dialysis section of the white blood cell; Fig. 60 is a schematic view showing a portion of the photodiode array of the photosensor; Fig. 61 is an enlarged view of the evaporator shown in the AP area of Fig. 55; Fig. 62 shows the PCR initiated by the linker; Figure 6 is a representative diagram of a test module with a lancet; Lutu 64 is a representative view of the structure of the LOC variant VII; Figure 65 is a plan view of the LOC variant VIII, the features and structures of all layers overlap each other Figure 66 is an enlarged view of the CA area shown in Figure 65; a partial perspective view of Figure 67 illustrates the layered structure of the LOC variant VIII in the CA area shown in Figure 05; Figure 68 is shown in Figure 66. Figure 69 is a representative view of the structure of the LOC variant XIV; Figure 71 is a representative representation of the structure of the LOC variant XLI: Figure 72 is a representation of the structure of the LOC variant XLI; Representative of the structure of the LOC variant XLII; Figure 73 is a representative representation of the structure of the LOC variant XLIII: Figure 74 is a representative representation of the structure of the LOC variant XLIV: Figure 75 is a representative representation of the structure of the LOC variant XLVII; 76 is a representative diagram of the structure of the LOC variant X; Fig. 77 is a perspective view of the LOC variant X; -314· 8 201211533 Fig. 78 is a LO A plan view of C variant X, which independently shows the structure of the CMOS + MST device; Figure 7 9 is a perspective view of the lower side of the upper disc, the reagent receptacles are indicated by dashed lines; Figure 80 is a plan view showing only the features of the upper cover independently; The 81 series shows a plan view in which all features overlap each other and indicates the position of the DA to DK zone;

82 is an enlarged view of the DA area shown in FIG. 81; FIG. 83 is an enlarged view of the DB area shown in FIG. 81; FIG. 84 is an enlarged view of the DC area shown in FIG. 81; An enlarged view of the DD area; Fig. 86 is an enlarged view of the DE area shown in Fig. 81; Fig. 87 is an enlarged view of the DF area shown in Fig. 81; Fig. 88 is an enlarged view of the DG area shown in Fig. 81; Fig. 81 is an enlarged view of the DH area shown in Fig. 81; Fig. 90 is an enlarged view of the DJ area shown in Fig. 81: Fig. 91 is an enlarged view of the DK area not shown in Fig. 81; Fig. 92 is a DL of Fig. 81 Magnified view of the area: Figure 93 is a circuit diagram of a differential imager; Figure 94 is a schematic illustration of a CMOS controlled flow rate sensor; Figure 95 illustrates the reaction occurring during the electrochemiluminescence (ECL) process; Figure 96 schematically illustrates three different Anode configuration: Figure 97 is a partial cross-sectional view of the anode and cathode in the hybridization chamber; Figure 98 is a schematic illustration of the anode surrounding the peripheral edge of the photodiode with a circular geometry - 315 - 201211533: Figure 99 is schematically illustrated around the photodiode A ring-shaped geometric anode within the edge; Figure 1 〇〇 schematically illustrates the anode of the series to increase the side edge of the Figure 101 illustrates the use of a transparent anode to maximize surface area coupling and ECL signal detection; Figure 102 illustrates the use of an anode fixed to the top of the hybrid chamber to maximize surface area coupling and EC L signal detection; Figure 103 schematically illustrates the cathode Figure 1 04 shows a test module and test module reader configured for ECL detection; Figure 1 05 is a diagram of electronic components in a test module configured for ECL detection Figure 1 06 shows the test module and the selective test module reader; Figure 1 07 shows the test module and test module reader and the host system that stores various databases; Figure 108A and 108B illustrate the aptamer and Protein binding to produce a detectable signal; Figures 109A and 109B illustrate the binding of two aptamers to a protein to produce a detectable signal; Figure 1 10A and 1 10B illustrate the binding of two antibodies to a protein to produce a detectable signal Figure 1 1 1 is a structural representation of the LOC variant L with ECL detection; -316- 201211533 Figure 1 is a perspective view of the L〇C variant L; Figure 113 is a plan view of the LOC variant L, which is independent Figure 1 is a bottom perspective view of the upper cover of the LOC variant L, the reagent reservoirs are shown in phantom; Figure 115 is a plan view of the LOC variant L, which separately shows the features of the upper cover; Figure 16 is a plan view showing that all features of the LOC variant L overlap each other and indicates the position of the GA to GL region; Figure 117 is an enlarged view of the GA region shown in Figure 116; Figure 118 is the GB shown in Figure 116 Figure 119 is an enlarged view of the GC region shown in Figure 116; Figure 120 is an enlarged view of the GD region shown in Figure 16; Figure 121 is an enlarged view of the GE region shown in Figure 116; Fig. I23 is an enlarged view of the GG area shown in Fig. 116: Fig. 124 is an enlarged view of the GH area shown in Fig. 116; Fig. 125 is shown in Fig. Figure 126 is an enlarged view of the GK area shown in Figure 16; Figure 127 is an enlarged view of the GL area shown in Figure 16; Figure 128 is a representative view of the LOC device with insulated grooves Figure 1 is a schematic diagram of an electrochemiluminescence resonance energy transfer probe in a closed configuration; Figure 130 is an electrochemiluminescence resonance energy transfer in an open hybrid configuration -31 7- 201211533 Probe pattern; Figure 1 3 1 is a diagram of the light-emitting linear probe connected to the primer during the first round of amplification; Figure 1 32 is a diagram of the light-emitting linear probe connected to the primer during the subsequent amplification period Figure 133A to 133F illustrate the heating cycle of the illuminating stem-loop probe connected to the primer; Figure 3-4 shows the stem-loop configuration of the negative control luminescent probe; Figure 1 3 5 schematically illustrates Figure 134 The open configuration of the negative control luminescent probe; Figure 136 schematically illustrates the stem-loop configuration of the positive control luminescent probe; Figure 137 illustrates the open configuration of the positive control luminescent probe of Figure 136; Figure 138 An enlarged view of the hybridization chamber of LOC variant L: Figure 139 is an enlarged view of the array of hybridization chambers of LOC variant L showing the distribution of such hybridization chambers. [Main component symbol description] I 〇: Test module II: Test module 1 2: Test module reader 13: Case 14: Micro USB connector 1 5: Inductor-318- 201211533 1 6 : Micro USB port 17 : Touch screen 1 8 : Display screen 19 : Button 20 : Start button 21 : Honeycomb radio 22 : Aseptic sealing tape φ 2 3 : Wireless network connection 24 : Large container 2 5 : Satellite navigation system 26 : Light-emitting diode 2 7 : Data storage 28 : Mobile phone / smart phone 2 9 : USB compatible LED driver 30 : Laboratory chip (LOC) device φ 3 1 : Power conditioner 3 2 : Capacitor 33 : Clock 34 : Controller 35: register 36: USB device driver 3 7 : drive 38 : RAM 3 9 : drive -319 - 201211533 40 : program and data flash memory 41 : register 42 : processor 43 : program memory 44 : Light sensor 45: indicator 46: upper cover

47 : USB Power / Indicator Module 48 : COMS + MST Chip 4 9 : Porous Element 5 2 : Hybridization and Detection Department 5 4 : Reservoir 5 6 : Reservoir 57 : Printed Circuit Board (PCB)

5 8 : receptacle 60 : reservoir 62 : reservoir 6 4 : lower sealing layer 66 : top layer 6 8 : sample inlet 70 : pathogen dialysis section 72 : waste channel 74 : target channel 76 : waste unit (reservoir) -320- 8 201211533 7 8 : Reservoir layer 80 : upper cover channel layer 8 2 : upper sealing layer 8 4 : 矽 substrate 86 : CMOS circuit 87 : MST layer 8 8 : passivation layer φ 90 : MST channel 92 : drop port 94 : Upper cover channel 96 : Riser 97 : Wall 98 : Meniscus anchor 1 00 : MST channel layer 101 : Laptop / notebook • 102 : Capillary start feature (CIF) 103 : Dedicated reader 1 〇5: Desktop computer 106: Boiling start valve 107: E-book reader 108: Boiling start valve 109: Tablet computer 1 1 〇: Hybridization room array η 1 : Epidemiological data host system -321 - 201211533 1 12 : Amplification unit 1 1 3 : Gene data host system 1 1 4 : Culture unit 1 15 : Electronic health record (EHR) host system 1 1 6 : Anticoagulant 1 1 8 : Surface tension valve 1 1 9 : Sample flow 1 2 0 : meniscus 121: electronic medical record (EMR) host system 1 2 2 : vent 123: personal health record (P HR) host system 124 : protein Phytobody detection chamber array 1 2 5 : Network 126 : Boiling start valve 1 2 8 : Surface tension valve 1 3 0 : Chemical lysis part 1 3 1 : Mixing part 1 3 2 : Surface tension valve 1 3 3 : Incubator Inlet channel 1 34 : Drop port 1 36 : Light window 1 38 : Surface tension valve 140 · Polymerase surface tension valve 141 : Aptamer - 322 - 201211533 142 : Specific protein 143 : Donor 144 : Recipient 145 : Antibody 146 : Valve inlet 147: Oligonucleotide 1 48: Valve outlet φ 149 : Connector 1 50 : Drop port 1 5 2 : Heater 1 5 3 : Valve heater contact 1 5 4 : Heater 1 5 6 : Heater Contact 158: Microchannel 160: Amplification section outlet channel φ 164: Hole array 165: Hole 166: Capillary initiation feature 168: Dialysis riser hole 170: Temperature sensor 174: Liquid sensor 175: Diffusion barrier 1 7 6 : Flow path 178 : End point liquid sensor - 323 201211533 1 80 : Hybridization chamber 1 8 2 : Heater 1 84 : Photodiode 1 85 : Active area 186 : Oligonucleotide FRET probe 187: Triggering photoelectric Diode 1 8 8 : Water reservoir 190 : Evaporator 1 9 1 : Heater 192 : Water supply channel 1 93 : Riser 1 94 : Descent 1 9 5 : Top metal layer 1 9 6 : Humidification 198: first rising hole 202: capillary starting feature 204: dialysis MST channel 206: valve 207: valve 208: liquid sensor 210: microchannel 212: intermediate MST channel 218: TiA1 electrode 220: TiA1 electrode -324 - 201211533 222: gap 23 2 : humidity sensor 2 3 4 : heater 237: ECL probe 23 8 : target nucleic acid sequence 240 : ring 242 : stem φ 248 : quencher 2 5 0 : fluorescence emission 28 8 : Sample input and preparation 2 90: Extraction 2 9 1 : Culture 2 9 2 : Amplification 29 3 : Before hybridization 294 : Detection • 296 : First electrode 29 8 : Second electrode 3 〇 0 : Preprogrammed delay 301 : Experiment Chamber wafer (LOC) device 3 2 8 : dialysis unit 3 7 6 : conductive column 3 7 8 : positive control probe 3 8 0 : negative control probe 3 8 2 : calibration chamber -325 201211533 3 9 0 : lancet 3 92 : lancet release button 4 0 8 : membrane seal 4 1 0 : membrane guard

5 1 8 : LOC variant VIII 5 94 : Interface layer 596 : Interface channel 598 : Interface channel 600 : Side channel 602 : Interface target channel 604 : Interface waste channel

673 : LOC variant XLIII

674 : LOC variant XLIV 677 : LOC variant XLVII 679 : LOC variant XLIX 6 8 2 : small component dialysis section 6 8 6 : dialysis section 694 : amplification blocker 696 : probe sequence 698 : sequence 700 : oligo Nucleotide primer 705: ECL probe 706: complementary sequence 708: stem stock-326- 201211533

7 1 0 : Another strand 728 : LOC change 729 : LOC change 73 0 : interface pass 7 3 2 : interface pass 734 : small component 7 3 6 : large component 73 8 : drop port 7 4 0 : flow rate sense 766 : Waste storage 7 6 8 : Waste storage 7 8 6 : Negative pair 7 8 7 : Positive pair 7 8 8 : Difference into 7 9 0 : Pixel 7 9 2 : Virtual image 794 : Read 795 : Read 797 : M4 801 : MD4

8 0 3 : Pixel power 805 : virtual image 8 0 7 : switch 809 : switch body X body L channel upper cover channel upper cover channel detector device ECL probe according to ECL probe image circuit element] J d container capacitor 201211533 81 1 : "Read - Line" switch 8 13 : Virtual "Read_Line" switch 8 1 4 : Bending heater element 8 1 5 : Switched capacitor amplifier 8 1 7 : Differential signal 8 60 : ECL excitation electrode 8 62: Light emission 8 64 : Luminous group 8 66 : Co-reactant 868 : Excited species 8 70 : ECL excitation electrode 872 : Solution 8 74 : ECL cell 8 7 6 : Dielectric gap 8 7 8 : Comb structure anode 8 8 0: finger-like structure 8 8 2 : curved configuration 8 84 : curve 8 8 6 : configuration 8 8 8 : tooth structure 8 9 0 : branch structure 892 : volume 8 94 : closed area 8 96 : groove - 328-

Claims (1)

201211533 VII. Patent application scope: 1. A microfluidic device for simultaneously detecting multiple states of a patient, the microfluidic device comprising: an inlet for receiving a sample of biological material extracted from the patient; a microsystem technology (MST) layer of the detection portion, the layer having a probe φ array for reacting with a target molecule in the sample to form a probe-target complex, the target molecule being in the medical state of the patient And a light sensor for detecting the probe-target complex; wherein the probe array has more than 1,000 probes. 2. The microfluidic device of claim 1, wherein the target molecular tag nucleic acid sequence and the probe array has a probe configured to hybridize to the target nucleic acid sequence to form a probe-target hybrid. 3. The microfluidic device of claim 1, wherein the target molecule is a protein and the probe array has a probe configured to hybridize or conjugate with the target protein to form a probe-target complex. . 4. The microfluidic device of claim 2, further comprising a CMOS circuit and a support substrate, the CMOS circuit being located between the MST layer and the support substrate, wherein the light sensor is inserted into the CMOS circuit Photodiode array. 5. The microfluidic device of claim 4, further comprising an array of hybridization chambers comprising probes, wherein each probe has a nucleic acid sequence complementary to one of the target nucleic acid sequences and electrochemiluminescence (ECL) illumination The group, and each -329-201211533 hybridization chamber has an electrode for generating an excited state of the ECL luminophore, wherein the ECL luminophore emits photons of light in an excited state. 6. The microfluidic device of claim 5, further comprising a nucleic acid amplification portion upstream of the hybridization chamber array, the nucleic acid amplification portion being configured to amplify the target nucleic acid sequence. 7. The microfluidic device of claim 6, wherein the nucleic acid amplification portion is used to amplify a polymerase chain reaction (PCR) portion of the target nucleic acid sequence before the target nucleic acid sequence hybridizes to the ECL probe. . 8. The microfluidic device of claim 1, wherein the MST layer has a plurality of MST channels configured to aspirate a fluid containing the target nucleic acid sequence by capillary action through the PCR portion and enter The hybridization chamber. 9. The microfluidic device of claim 8, wherein the CMOS circuit has a pad for electrically connecting to an external device, wherein the CMOS circuit is configured to convert an output from the photodiode to display the A signal that the ECL probe hybridizes to the target nucleic acid sequence and provides the signal to a pad for transmission to the external device. 10. The microfluidic device of claim 9, wherein each of the probes has a functional portion of photon emission that is quenched by resonance energy transfer to quench the ECL luminophore. The microfluidic device of claim 9, wherein the probe is configured to quench photon emission of the ECL luminophore when the probe forms a probe-target hybrid. The functional moiety is further derived from the ECL luminophore. The microfluidic device of claim 5, wherein the CMOS circuit is configured to provide an electrical pulse to the electrode, the electrical pulse period being less than 6.9 seconds. 13. The microfluidic device of claim 12, wherein the electrical current of the electrical pulse is between 0.1 nanoamperes and 69.0 nanoamperes. The microfluidic device of claim 5, wherein the CMOS circuit is configured to control the temperature of the hybridization chamber during the hybridization of the probe to the target nucleic acid sequence. The microfluidic device of clause 14, further comprising a hybrid heater controlled by the CMOS circuit to provide thermal energy for hybridization. 16. The microfluidic device of claim 15 further comprising a fluid flow path from the PCR portion to the end liquid sensor, the hybrid chamber being disposed along two sides of the fluid flow path. The microfluidic device of claim 6, wherein the flow Φ body flow path is configured to draw fluid from the PCR portion to the liquid end point sensor by capillary action and each of the hybrid chambers The fluid from the fluid flow path is configured to be filled by capillary action such that during use, the CMOS circuit reacts to activate the hybridization heater from the liquid endpoint sensor indicating that the fluid has reached the output of the liquid endpoint sensor. 18. The microfluidic device of claim 7, wherein each of the hybridization chambers has a volume of less than 9, 〇〇〇 cubic micrometers. The microfluidic device of claim 5, wherein the photodiode is less than L600 microns from the ECL probe. The microfluidic device of claim 16, further comprising a plurality of reagent reservoirs for treating different reagents required for the fluid, wherein the flow system is drawn from the inlet by capillary action to The endpoint sensor does not require the addition of liquid from a source other than the microfluidic device. -332-