CN211586663U - A biological detection chip - Google Patents

Abstract

The utility model discloses a biological detection chip, including the base member, be provided with on the base member by biological detection chip's rotation center along the radial outside preceding processing chamber, distribution chamber, water conservancy diversion chamber and the reaction chamber that communicates in order, the intercommunication has the voltage limiting return bend between water conservancy diversion chamber and the reaction chamber, and two inboards of the upper end entry in water conservancy diversion chamber have designed a binding off structure to the inboard extension respectively, are equipped with the sample addition mouth on the preceding processing chamber, still are equipped with the exhaust hole of its inside each cavity of intercommunication and external environment on the base member. The biological detection chip has the pressure-limiting bent pipe with the function of the flow control valve, the liquid can be transferred step by step only by controlling the centrifugal rotating speed, an additional fluid control mechanism is not needed, the operation process is simple and easy, manual operation intervention is not needed in the middle process, the workload of testers is reduced, and the detection efficiency is improved.

Description

A biological detection chip

technical field

The utility model relates to the technical field of biological detection supporting equipment, in particular to a biological detection chip.

Background technique

At this stage, microfluidic chips are the hot spot in the development of the current micro-total analysis system. It takes the chip as the operating platform, and combines with biology, chemistry, drug screening and other technologies to complete reagent loading, separation, reaction, and detection. The whole process including the device. In recent years, with the rapid development of microfluidic technology, microfluidic chips have played an increasingly important role in the fields of life sciences, analytical chemistry and biomedicine due to their advantages of integrated miniaturization and automation, small volume of reagents, and high throughput. The increasingly important role is not only for the field of routine biological detection, but also for other related fields with special operating environment or operational requirements.

Taking aerospace biomedicine as an example, space microbes are a major safety issue facing long-term manned spaceflight, which seriously threatens the life and health of astronauts and the long-term safe operation of spacecraft. The growth of microorganisms in manned spacecraft will pollute the environment, cause astronauts to become infected or sick, corrode materials, and cause equipment failure. If microorganisms that mutate in space are brought back to the earth, they will also threaten the ecological security of the earth. Therefore, the development of advanced rapid detection technology for potential pathogenic microorganisms in the space station can help to quickly identify the cause when personnel infection and environmental abnormalities occur, and formulate targeted measures for effective intervention, which will help ensure the physical and mental health of astronauts and spaceflight during orbital flight. The smooth execution of the task is of great significance and value. The routine laboratory microbial nucleic acid detection technical process includes sample processing and nucleic acid extraction, nucleic acid amplification, nucleic acid detection and result analysis, etc., requiring professionals to be equipped with certain safety protection in professional laboratories and pipetting on different professional instruments, etc. Operation, the space station cannot carry out conventional liquid operations due to special environmental requirements such as aerospace microgravity, low power consumption, weight and volume.

At present, it is not easy for microfluidic chips to achieve a high degree of integration and less external driving force. Most of the integration requires a lot of extra micro-pumps and micro-valves. The equipment and devices required to drive these pumps and valves are also relatively complex, adding a micro-fluidic chip. complexity, manufacturing difficulty, cost, and reduced reliability. For example, CN203750554U discloses a microfluidic chip for multi-index detection, but the chip is also limited to only provide a detection platform, and other auxiliary processes such as other instruments are required to complete the process of sample pretreatment; CN205797240U discloses a It is a fully integrated microfluidic chip for multi-index detection of microfluidics, but the chip has an additional mixing valve structure, and additional auxiliary equipment is required to accurately operate the structure to complete the whole process reaction.

In addition, for general clinical testing, or related testing in other biochemical fields, due to the limitations of current technological development, the integration of chips and related equipment required for testing and testing is low, and it is usually only able to complete the final Reaction operations, many pre- or main inspection and detection operations need to rely on a large number of manual operations to complete, and the packaging process requirements of related equipment are relatively high, and some more special equipment even needs to rely on additional fluid control structures to achieve the detection and testing. Reasonable control not only restricts the operation efficiency of the entire detection step, but also limits the application environment of the relevant biochips in the inspection and detection process, and also adversely affects the accurate and efficient operation of the relevant inspection and detection.

Therefore, how to provide a biological detection chip to reduce manual operations, get rid of the dependence on the fluid control structure, improve the detection efficiency, and expand the scope of application is a technical problem that needs to be solved by those skilled in the art.

Utility model content

The purpose of the present utility model is to provide a biological detection chip, the relevant operation and use process of the biological detection chip is simple and easy, without the need for an additional fluid control structure, with high detection efficiency and a wide range of applications, which can be used for biological organisms in aerospace microgravity environments. detection test.

In order to achieve the above purpose, the present invention provides a biological detection chip, which includes a base body, and the base body is provided with a pretreatment cavity, a distribution cavity, a distribution cavity, a pretreatment cavity, a distribution cavity, a A diversion cavity and a reaction cavity, the diversion cavity and the reaction cavity are connected with a pressure-limited elbow, the two inner sides of the upper inlet of the diversion cavity are respectively designed with a closing structure extending inwardly, the front The processing chamber is provided with a sample injection port, and the base body is also provided with an exhaust hole that communicates with the internal chambers and the external environment.

Preferably, the length of the closing structure is less than half of the width of the diversion cavity.

Preferably, the guide cavities are sequentially arranged along the circumferential direction of the centrifugal rotation circle of the biological detection chip or along the extension direction of the involute of the centrifugal rotation circle of the biological detection chip.

Preferably, along the radial direction of the centrifugal rotation circumference of the biological detection chip, a sedimentation chamber is connected downstream of the reaction chamber, and the near-rotation center end of the sedimentation chamber is connected to the far-rotation center end of the reaction chamber.

Preferably, the middle part of the pressure-limiting elbow is a U-shaped pipe section, two ends of the U-shaped pipe section are respectively connected with straight pipe sections extending along the flow direction of the diversion cavity, and the main body of the U-shaped pipe section is straight The pipe is inclined relative to the straight pipe section.

Preferably, a buffer cavity is communicated between the diversion cavity and the reaction cavity, the pressure-limiting elbow is communicated between the buffer cavity and the diversion cavity, and the buffer cavity is connected to the reaction cavity. A capillary is communicated between the cavities.

Preferably, the distribution cavity is communicated with a plurality of the diversion cavities, each of the diversion cavities is communicated with a buffer cavity through one of the pressure-limiting elbows, and each of the buffer cavities is communicated with a buffer cavity. The capillary is communicated with one of the reaction chambers.

Preferably, an upstream conduit is communicated between the feed end of the distribution cavity and the pretreatment cavity, and a large-diameter buffer cavity is provided in the middle of the upstream conduit.

Preferably, a downstream conduit is communicated between the near-rotation center end of the distribution chamber and the near-rotation center end of the pretreatment chamber, and the exhaust hole is provided on the downstream conduit.

Preferably, a waste liquid chamber is communicated with the end of the distribution chamber.

Preferably, the base body is provided with a plurality of the pre-processing chambers, and each of the pre-processing chambers is sequentially arranged and communicated radially outward from the rotation center of the biological detection chip.

The working process of the biological detection chip provided by the present invention is as follows:

The relevant reagents and test samples required for the test and detection are added into the pretreatment chamber through the injection port, and the air in the pretreatment chamber is discharged to the external environment through the exhaust hole, and then the injection port is closed. Then put the chip into the auxiliary control device for heating and temperature control or light treatment. After the treatment is completed, the centrifugal rotating device is used to drive the biodetection chip to rotate as a whole, so that the liquid in the pretreatment chamber flows into the distribution chamber under the action of centrifugal force, and enters respectively. In each diversion cavity, at the same time, the gas in the distribution cavity and the diversion cavity is squeezed through the downstream pipeline into the pretreatment cavity to achieve air pressure balance. When the liquid reaches the diversion cavity at a low flow rate, although the diversion cavity and the reaction cavity are also in a connected state, because the pressure limiting elbow relies on its own conduction threshold to form a valve effect, the liquid pressure is smaller than the valve threshold at this time. Therefore, it can prevent the liquid from entering the reaction chamber through the pressure-limiting elbow in the non-test state. The closed structure of the guide cavity can prevent the reagents stored in the adjacent guide cavity from interfering with each other during the mixing process. When the rotational speed of the centrifugal rotating device is increased, and the liquid pressure in the diversion chamber is greater than the conduction threshold of the pressure-limiting elbow, the liquid can enter the reaction chamber for reaction, so as to complete the relevant biological detection operations. The precipitation chamber is used to collect the solids that appear after the reaction in the upstream reaction chamber 12 .

The utility model has the following beneficial effects:

1) The biodetection chip has a pressure-limiting elbow that acts as a flow control valve, and the liquid can be transferred step by step only by controlling the centrifugal speed without the aid of an additional fluid control mechanism. The operation process is simple and easy, and no intermediate process is required. Manual operation intervention reduces the workload of the test personnel and improves the detection efficiency;

2), the chip integration is high, the structure is simple, and the manufacturing and packaging costs are low;

3) The chip detection results are accurate and reliable, and the operation process is precise and controllable. It can be widely used in automatic nucleic acid extraction and multiple amplification detection of viruses or cells in various sample types such as saliva, throat swabs, and cervical swabs. It works normally in the ground environment and can also be applied to microgravity environments such as space stations, which greatly expands the scope of application of the product.

Description of drawings

In order to more clearly illustrate the embodiments of the present utility model or the technical solutions in the prior art, the following briefly introduces the accompanying drawings that need to be used in the description of the embodiments or the prior art. Obviously, the accompanying drawings in the following description These are just some embodiments of the present invention, and for those of ordinary skill in the art, other drawings can also be obtained from these drawings without creative effort.

1 is a schematic structural diagram of a biological detection chip in the first embodiment of the present invention;

2 is a schematic structural diagram of a biological detection chip in a second embodiment of the present invention;

3 is a schematic structural diagram of another biological detection chip in the first embodiment of the present invention;

4 is a schematic structural diagram of a biological detection chip in a fourth embodiment of the present invention;

5 is a cross-sectional view of an internal structure of a biological detection chip provided by the present invention;

6 is a cross-sectional view of another internal structure of the biological detection chip provided by the utility model;

7 is a schematic structural diagram of a biological detection chip with two substrates on one substrate in the present invention;

8 is a schematic structural diagram of a biological detection chip with three substrates disposed on one substrate in the present invention;

FIG. 9 is a schematic diagram of the diversion cavity and its closing structure of part A in FIG. 8;

10 is a schematic structural diagram of a biological detection chip with four substrates on one substrate in the present invention.

In Figures 1 to 10:

101-substrate, 102-package board, 11-base body, 110-centrifugal rotation circle, 111-pretreatment chamber, 112-distribution chamber, 113-direction chamber, 114-sample inlet, 115-upstream conduit, 116- Downstream conduit, 117-large diameter buffer chamber, 118-waste chamber, 119-decontamination chamber, 12-reaction chamber, 121-buffer chamber, 122-pressure limiting elbow, 123-capillary, 124-precipitation chamber, 13- Exhaust hole, 14-cuff structure.

Detailed ways

The core of the utility model is to provide a biological detection chip, the related operation and use process of the biological detection chip is simple and easy, no additional fluid control structure is needed, the detection efficiency is high, and the application range is wide.

In order to make those skilled in the art better understand the solution of the present invention, the present invention will be further described in detail below with reference to the accompanying drawings and specific embodiments.

Please refer to FIGS. 1 to 10, wherein, FIGS. 1 to 4 respectively show the structures of the biological detection chips of the three embodiments; FIGS. 5 and 6 respectively show cross-sectional views of the internal structures of the two biological detection chips; 7. FIG. 8 and FIG. 10 respectively show a schematic structural diagram of a biological detection chip with two substrates, three substrates and four substrates respectively provided on one substrate.

The utility model provides a biological detection chip, which includes a base body 11, and the base body 11 is provided with a pre-processing cavity 111, a distribution cavity 112, a guide cavity 113 and In the reaction chamber 12, a limited-pressure elbow 122 is communicated between the diversion cavity 113 and the reaction cavity 12. The two inner sides of the upper inlet of the diversion cavity 113 are respectively designed with a closing structure 14 extending inwardly, and the pretreatment cavity 111 is provided with The sample introduction port 114 and the base body 11 are also provided with an exhaust hole 13 that communicates with the internal chambers and the external environment.

It should be noted that, in the biological detection chip provided by this solution, the substrate 11 and its respective chambers are used as a complete integrated reaction unit. 1, 2, 3, 4 or more substrates 11, the integrated reaction unit represented by each substrate 11 can independently complete a biological detection test, as shown in Figure 7, Figure 8 and Figure 10. A biological detection chip structure in which two substrates 11 , three substrates 11 and four substrates 11 are respectively provided on a substrate 101 is shown. The overall structure of the substrate 101 may be a circular center plate with a central hole, or be designed to be fan-shaped, square, oval or other structures. The circumferential directions of the centrifugal rotation circles are arranged in sequence.

It should be noted that, when the above-mentioned substrate 101 has a circular plate structure, its diameter is in the range of 40 to 600 mm, preferably 60 to 150 mm in diameter. The material of the substrate 101 is a mixture of one or more of glass, silicon, metal, and polymers. In principle, any material that can meet the actual use requirements of the biological detection chip can be used.

In this solution, biological or chemical reagents for sample pretreatment can be stored in the pretreatment chamber 111 in advance. The storage form of the reagents can be liquid or solid, and can be droplet, paste, gel, film, or powder. etc., which can be one substance or a mixture of several substances, and can be placed in one or more places in the pre-processing chamber 111 . Preferably, the storage reagent contains components that can be used for sample liquefaction and cell and/or pathogen lysis, and can complete the lysis of cells and/or pathogens contained in the sample under certain conditions (such as 37°C to 95°C). Cracking. The reagent components may include but are not limited to sodium hydroxide, sodium bisulfate, DTT, TCEP, guanidine isothiocyanate, guanidine hydrochloride, SDS, TritonX-100, Tween-20, CTAB, proteinase k, lysozyme and other components One or more of these ingredients, and several auxiliary ingredients, such as one or more of excipients, preservatives and other ingredients.

In this solution, biological or chemical reagents for mixing or reacting with the pretreated samples can be stored in the diversion cavity 113 in advance, and the storage forms of the reagents can be drop, paste, gel, film, In powder form, it can be one substance or a mixture of several substances. Preferably, the reagent contains chemical components that can be used in nucleic acid amplification reactions, including but not limited to DNA polymerase, RNA polymerase, reverse transcriptase, recombinase, nickase, nucleic acid repair enzyme, restriction endonuclease One or more of enzymes, magnesium ions, potassium ions, dNTPs, rNTPs, PEG (400-20000), BSA, TE, betaine and other components, as well as several auxiliary components, such as fluorescent dyes, excipients, preservatives one or more of the ingredients.

In this solution, biological or chemical reagents for mixing or reacting with previously processed samples can be stored in the reaction chamber 12 in advance, and the storage forms of the reagents can be drop, paste, gel, film, or powder. , which can be one substance or a mixture of several substances. Preferably, the reagent contains components that can be used for specific nucleic acid amplification reaction or result indication, including one or more groups of oligonucleotides with specific sequences or specific fluorescent labels that can specifically bind to a certain DNA template fragments, and several auxiliary components, such as one or more of fluorescent dyes, excipients, preservatives and other components.

It should be pointed out that, in actual operation, the above-mentioned reagents can be stored by adding liquid or solid substances directly into each chamber, or drying and solidifying by natural drying, drying, air-drying, freeze-drying, etc. after adding liquid substances.

Preferably, along the radial direction of the centrifugal rotation circle of the biological detection chip, a sedimentation chamber 124 is connected downstream of the reaction chamber 12 , and the near-rotation center end of the sedimentation chamber 124 is connected to the far-rotation center end of the reaction chamber 12 . A precipitation chamber 124 is communicated downstream of the reaction chamber 12 for collecting the solids that appear after the reaction in the upstream reaction chamber 12, so as to collect and process centrally and prevent these solids from interfering with subsequent related reactions.

It should be noted that the near-rotation center end described in this article refers to the end position of the component that is closest to the rotation center of the biological detection chip, and the far rotation center end refers to the component that is farthest from the rotation center of the biological detection chip. end position.

It should be noted that the shape of the pressure limiting elbow 122 in this solution can be a circular arc or a smooth curved pipe, or a zigzag pipe, including but not limited to U-shaped pipe, S-shaped pipe, Ω-shaped pipe, L-shaped tube, etc. Preferably, in this solution, the middle of the pressure-limiting elbow 122 is a U-shaped pipe section, and the two ends of the U-shaped pipe section are respectively connected with straight pipe sections extending along the diversion direction of the diversion cavity 113 . The pipe sections are relatively inclined. The diversion direction of the diversion cavity 113 is the direction in which the diversion cavity 113 guides the liquid flow along the radial direction of the centrifugal rotation circle of the biological detection chip. Side straight tube.

The diameter of the pressure-limiting elbow 122 is 0.1 to 1 mm, and the preferred size is 0.2 mm.

Preferably, a buffer cavity 121 is communicated between the diversion cavity 113 and the reaction cavity 12 , the pressure limiting elbow 122 is communicated between the buffer cavity 121 and the diversion cavity 113 , and a capillary is communicated between the buffer cavity 121 and the reaction cavity 12 123. The pressure-limiting elbow 122 can cooperate with the capillary 123 to effectively block and restrict the flow of reagents and samples between the diversion chamber 113 and the reaction chamber 12, thereby further improving the operation accuracy and controllability of the relevant detection reaction, while the buffer chamber 121 can effectively buffer the liquid passing through the diversion cavity 113, so as to avoid the high-speed flowing high-pressure liquid causing structural impact on the main structure of the reaction cavity 12 or other phenomena that may adversely affect the reaction process and detection results. To further ensure the reliability and accuracy of the test results.

It should be further explained that, in practical applications, the purpose of adjusting the feed conduction threshold of the corresponding reaction chamber 12 can be achieved by adjusting and replacing the pressure-limiting elbow 122 and the capillary tube 123 of different sizes and specifications, so as to meet the requirements for different conditions under different working conditions. The test materials are subjected to corresponding testing tests.

Preferably, the distribution chamber 112 is connected with a plurality of guide chambers 113 , each guide chamber 113 is connected with a buffer chamber 121 through a pressure limiting elbow 122 , and each buffer chamber 121 is connected with a reaction chamber through a capillary 123 . Cavity 12. The biological detection chips shown in FIG. 1 to FIG. 4 each have 10 guide cavities 113 and 10 corresponding reaction cavities 12 .

It should be noted that the diversion chamber 113 in this solution can pre-store reagents, thereby structurally replacing the mixing chamber of the chip in the prior art, thereby simplifying the chip structure, and the pre-stored reagent concentrations in the plurality of diversion cavities 113 can be Consistent, may not be the same. The diversion cavity 113 not only has the function of quantifying the liquid, but also quantifies the liquid volume more accurately. Moreover, the liquid can be mixed with the pre-stored reagents in the diversion cavity 113 when the liquid is distributed into the diversion cavity 113, and the mixing effect is better than the existing one. In the technology, it is better to concentrate on mixing in one chamber.

It should be noted that the shape of the guide cavity 113 may be a square, a rectangle, a triangle, a rounded rectangle, a semicircle, a semiellipse, etc., or a combination of the above-mentioned shapes. The shape of the guide cavity 113 can be the same or different, and its area or volume can be the same or different, so as to achieve the purpose of uniform or non-uniform distribution and transfer of liquid. The upper opening of the diversion cavity 113 is preferably designed to be closed. As shown in FIG. 8 and FIG. 9 , two inner sides of the upper inlet of the diversion cavity 113 are respectively designed with a closure structure 14 extending inwardly. Further preferably, the closure structure 14 The length of the guide cavity 113 is less than half of the width of the guide cavity 113. By designing the closing structure 14, it can be ensured that each adjacent guide cavity 113 can be avoided during the process of the liquid flowing into the guide cavity 113 and the process of mixing the liquid in the guide cavity 113. interfere with each other.

Preferably, the guide cavities 113 are sequentially arranged along the circumferential direction of the centrifugal rotation circle of the biodetection chip or along the extension direction of the involute of the centrifugal rotation circle of the biodetection chip, as shown in FIG. 1 , FIG. 2 and FIG. 7 . The centrifugation of the biological detection chip rotates the circular line 110 . When the diversion cavities 113 are arranged along the involute, the liquid can be distributed into each diversion cavity 113 more smoothly and evenly, that is, the diversion cavity 113 away from the upstream conduit 115 can be more fully distributed dispensed into the liquid.

It should be noted that, between the distribution chamber 112 and the pretreatment chamber 111 , the end-to-end connection can be performed only by means of connecting pipes, or the circulation connection can be formed by relying on upstream and downstream pipes.

Referring to FIG. 1 , in an embodiment, preferably, an upstream conduit 115 is communicated between the feed end of the distribution cavity 112 and the pretreatment cavity 111 , and a large diameter buffer cavity 117 is provided in the middle of the upstream conduit 115 . Specifically, the proximal rotating central end of the upstream conduit 115 is connected to the far rotating central end of the pretreatment chamber 111 , so as to transfer all the liquid in the pretreatment chamber 111 ; the distal rotating central end of the upstream conduit 115 is connected to the The feed end, that is, the end of the distribution chamber 112 near the center of rotation. The inner diameter of the large-diameter buffer chamber 117 is larger than the inner diameter of the upstream conduit 115 , and the large-diameter buffer chamber 117 can moderately buffer and divert the liquid flow from the pre-treatment chamber 111 into the upstream conduit 115 to stabilize the liquid flow. The pre-processing chamber 111 can be stably and efficiently passed into the downstream distribution chamber 112 and other related chambers.

Referring to FIG. 1 , in the above-mentioned embodiment, preferably, a downstream conduit 116 is communicated between the near-rotation center end of the distribution chamber 112 and the near-rotation center end of the pretreatment chamber 111 , and the exhaust hole 13 is arranged in the downstream conduit 116 on. The vent hole 13 is preferably provided at the proximal rotational center end of the downstream conduit 116 . During the sample adding process, the role of the downstream conduit 116 is to discharge the gas in the pretreatment chamber 111, the distribution chamber 112 and other related chambers to the external environment through the exhaust hole 13; during the centrifugal rotation process, the downstream conduit 116 and the The upstream conduit 115 forms a complete air circuit to ensure the smooth progress of the centrifugal rotation.

As shown in FIG. 1 , after the sample is added to the pretreatment chamber 111 through the sample injection port 114 , the air in the pretreatment chamber 111 reaches the downstream conduit 116 through the buffer pools and channels connected in parallel, as well as the upstream conduit 115 and the distribution chamber 112 . , and finally discharged from the exhaust hole 13 , and then the sample injection port 114 is closed with a film-like material with a certain viscosity. Centrifugal rotating equipment such as a rotary motor is used to provide centrifugal rotation driving force, and the liquid in the pre-processing chamber 111 is driven into the distribution chamber 112. At this time, the liquid in the pre-processing chamber 111 is driven by the low-speed centrifugal force, breaking through the large-diameter buffer chamber The blocking effect of 117 reaches distribution chamber 112 .

Please refer to FIG. 3 , which is a schematic structural diagram of another biological detection chip in the first embodiment of the present invention. Comparing the structure of the chip shown in FIG. 3 with the chip shown in FIG. 1 , the difference is that no closing structures are designed on both sides of the upper inlet of the diversion cavity 113 , and the structures of other components are the same as those shown in FIG. 1 .

Referring to FIG. 2 , in another embodiment, the downstream conduit 116 is only connected to the end of the distribution cavity 112 near the center of rotation and extends radially toward the center of rotation of the biological detection chip, and the end of the downstream conduit 116 is opened near the center of rotation There are exhaust holes 13 for exhausting the gas in the distribution chamber 112 and related chambers.

As shown in FIG. 2 , after the sample is added to the preprocessing chamber 111 through the sample injection port 114, the liquid in the preprocessing chamber 111 is driven and transferred to the distribution chamber 112 by using a matching device. The space occupied by the liquid in the processing chamber 111 begins to decrease, and its internal air pressure begins to decrease. At the same time, the liquid in the distribution chamber 112 begins to increase, and its internal air pressure begins to increase. During this process, although there is no downstream conduit 116 as shown in FIG. The increased air pressure in the distribution chamber 112 can be balanced by the upstream conduit 115 and the reduced air pressure in the pretreatment chamber 111. At this time, the liquid in the distribution chamber 112 increases and the liquid in the pretreatment chamber 111 decreases, and a pressure balance is achieved between the two. state. Since the supply of the driving force is continuous, the above-mentioned equilibrium state is also constantly changing, until the liquid in the pretreatment chamber 111 is completely transferred to the distribution chamber 112, and the final equilibrium is reached. It is easy to understand that the magnitude of the liquid driving force is negatively related to the width of the upstream conduit 112 . According to the above, as long as the driving force is less than the threshold value of the valve formed by the pressure limiting elbow 122 , the liquid can still be distributed in the diversion chamber 113 and not enter the reaction chamber 12 in advance. A downstream conduit 116 and an exhaust hole 13 may also be provided on the side of the distribution chamber 112 close to the rotation center, which is more conducive to transferring all the liquid in the pretreatment chamber 111 to the distribution chamber 112 .

Preferably, the inner surfaces of the pre-processing chamber 111 , the large-diameter buffer chamber 117 , the distribution chamber 112 , the guide chamber 113 , and the inner surface of the reaction chamber 12 may be physically or partially treated by physical or chemical treatment to make them more denser than the original surface of the substrate material. Hydrophobic, that is, the surface hydrophobization treatment. Preferably, the contact angle between the surface and the contained solution after hydrophobic treatment is 90°˜140°. Preferably, in this solution, surface hydrophilization treatment can also be pre-applied to the inner walls of other chambers and pipelines other than the above-mentioned chambers as a whole or in part, so as to meet the relevant detection and test requirements of different samples or different application environments.

It should be specially pointed out that, in practical applications, the above-mentioned reaction chambers 12 are preferably evenly distributed with equal diameters and equal distances along the circumferential extension direction of the centrifugal rotation circle of the biological detection chip, that is, two adjacent reaction chambers 12 and centrifugal The distance between the centers of rotation of the rotating equipment is equal, and the central angles corresponding to the arc segments between the two adjacent reaction chambers 12 are equal. This uniform distribution structure can effectively ensure that the reaction rates of the samples and reagents in each reaction chamber 12 are consistent and the reaction rate is guaranteed. fully.

Preferably, a waste liquid chamber 118 is communicated with the end of the distribution chamber 112 . The waste liquid generated during the relevant detection test can be passed into the waste liquid chamber 118 through the distribution chamber 112 under the action of centrifugal force for centralized collection. The waste liquid cavity 118 is also designed with a guide cavity 113 correspondingly, and the ratio of the volume of the guide cavity 113 of the waste liquid cavity 118 to the corresponding flow guide cavity 113 of the reaction chamber 12 is greater than 2, so as to ensure that there is enough volume to accommodate the excess liquid. In practical application, waste liquid chambers 118 may be provided at both ends of the distribution chamber 112 to ensure the waste liquid collection efficiency. The waste liquid chamber 118 may also be provided only at one end of the distribution chamber 112 . Of course, if multiple waste liquid cavities 118 are provided, it should be ensured that each waste liquid cavity 118 and each diversion cavity 113 are also arranged along the same arc extending direction to ensure the liquid diversion effect.

The waste liquid chamber 118 has various shapes, such as square, rectangle, triangle, rounded rectangle, semi-circle, semi-ellipse, etc., and can also be a combination of the above-mentioned shapes. In this solution, a pressure limiting elbow 122 can also be added between the waste liquid chamber 118 and the diversion chamber 113 to prevent the waste liquid from flowing back. When both ends of the distribution chamber 112 are provided with waste liquid chambers, the waste liquid chamber at one end of the feed port of the distribution chamber 112 is the impurity removal chamber 119 , and the volume of the diversion chamber 113 corresponding to the impurity removal chamber 119 is smaller than that of the reaction chamber 119 . The diversion cavity 113 corresponding to the cavity 12, and the volume of the impurity removal cavity 119 is larger than the volume of the diversion cavity 113 corresponding to the reaction cavity 12. The purpose of this setting is to allow the pretreatment cavity 111 to be removed during the initial low-speed centrifugal rotation. A section of liquid with impurities is first collected into the guide chamber 113 corresponding to the impurity removal chamber 119 to ensure that the effective sample reagent can enter the guide chamber 113 corresponding to each reaction chamber 12. The miscellaneous cavity 119 is then used to collect the liquid remaining in the pretreatment cavity 111 and the upstream conduit 115 .

In this solution, a pressure-limiting elbow 122 and other pressure-limiting communication parts can also be set between the waste liquid cavity 118 and the diversion cavity 113 and between the impurity removal cavity 119 and the diversion cavity 113, so as to reasonably control the conduction of the waste liquid cavity 118. The threshold value can prevent reagents or samples that should be involved in the relevant biochemical reactions of the detection test from entering the waste liquid chamber 118 by mistake, and also prevent the waste liquid from flowing back into the distribution chamber 112 .

Referring to FIG. 4 , in the third embodiment, preferably, a plurality of pre-processing chambers 111 are provided on the base body 11 , and each pre-processing chamber 111 is sequentially arranged radially outward from the rotation center of the biological detection chip and connect. In this way, multiple pre-processing steps of the sample can be realized, which can meet the needs of some biological testing and detection tests that require multiple pre-processing, such as multiple types of reagents or multiple types of samples, so as to further improve the test and detection efficiency of the biological detection chip provided by this scheme. It is extremely suitable. scope. Of course, each of the pre-processing chambers 111 can also be connected in parallel. During the centrifugal rotation, each of the pre-processing chambers 111 transfers liquid to the distribution chamber 112 at the same time. This arrangement can make the liquid in the pre-processing chamber 111 transfer to the distribution chamber 112 more quickly. , improve the detection efficiency.

It should be specially pointed out that, for different test requirements, the precipitation chamber 124 as described above may or may not be provided at the end of the reaction chamber 12, but whether or not the precipitation chamber 124 is provided, it should be used in practical applications. It is ensured that the volume of the diversion chamber 113 is not greater than the sum of the volumes of the downstream chambers corresponding to the precipitation chamber 124, so as to ensure that the biochemical reactions in the reaction chambers 12 are independent of each other and do not interfere with each other. Specifically, if the end of the reaction chamber 12 is provided with a sedimentation chamber 124, it should be ensured that the volume of the diversion chamber 113 is not greater than the sum of the corresponding volumes of the pressure limiting elbow 122, the reaction chamber 12 and the sedimentation chamber 124; If the precipitation chamber 124 is not provided at the end of the reaction chamber 12, it should be ensured that the volume of the diversion chamber 113 is not greater than the sum of the corresponding volume of the pressure-limiting elbow 122 and the reaction chamber 12; In all cases, it should preferably be ensured that the volume of the diversion cavity 113 is larger than the volume of the corresponding reaction cavity 12, so that the reaction is more sufficient, the reflection effect is better, and the detection result is more accurate and reliable.

It should be noted that the pre-processing chamber 111 , the distribution chamber 112 , the reaction chamber 12 and their subsidiary structures (buffer chamber 121 , precipitation chamber 124 , pressure-limiting elbow 122 , etc.) of the substrate 11 may be located on both sides of the substrate 101 , and The communication is carried out by means of small holes penetrating through the substrate 101 . The main part of the biological detection chip in this solution is the substrate 101 , the two sides of the substrate 101 are covered with the packaging board 102 , the pre-processing chamber 111 , the distribution chamber 112 , the diversion chamber 113 , the reaction chamber 12 , and the pressure-limiting elbow 122 are all located on the substrate 101 Between it and the packaging board 102 , the sample adding port 114 and the exhaust hole 13 are located on the packaging board 102 . The inter-board cavity structure formed by using the gap between the substrate 101 and the packaging board 102 can further improve the structural integration of the biological detection chip, optimize the airtightness and conduction efficiency of the corresponding internal cavity structure, so that the relevant biological The detection reaction is more sufficient and efficient.

It should be noted that, during actual processing and manufacturing, the substrate 101 and the packaging board 102 can be packaged and fastened by any method such as hot pressing, gluing, laser welding, ultrasonic welding or screw fastening, so as to ensure the overall assembly of the assembly. Sealing effect and assembly strength.

Please focus on Figure 5. In one embodiment, the preprocessing chamber 111 , the reaction chamber 12 and the pressure limiting elbow 122 are all located on the same side of the substrate 101 , and the distribution chamber 112 and the guide chamber 113 are located on the other side of the substrate 101 . The dislocation arrangement of different planes can make the main reaction chamber composed of the pretreatment chamber 111 , the reaction chamber 12 and the pressure-limiting elbow 122 relatively independent from the main guiding chamber composed of the distribution chamber 112 and the guiding chamber 113 They are separately packaged, thereby further improving the structural airtightness and structural integration of the internal chambers of the biological detection chip, and the related chambers located on different sides of the substrate 101 Ensure smooth flow of reagents and samples.

Please focus on Figure 6. In another embodiment, both sides of the substrate 101 are respectively provided with a pre-processing chamber 111 , a distribution chamber 112 and a guide chamber 113 , and one side of the substrate 101 is provided with a limiter communicating with each guide chamber 113 respectively. The bending tube 122 and the reaction chamber 12 are pressed, and there are guide holes penetrating the substrate 101 to communicate the chambers on both sides. This kind of main reaction chamber composed of a pre-processing chamber 111, a reaction chamber 12 and a pressure-limiting elbow 122 is provided on both sides of the substrate 101, and the above two sets of main reaction chambers are respectively connected with the same downstream set by the distribution chamber 112 and the pressure-limiting elbow. The main guide chambers formed by the guide chambers 113 are connected, so that in practical applications, two different samples or reagents can be placed in the two sets of pretreatment chambers 111 located on both sides of the substrate 101 respectively, and the corresponding In the processing process, after the pre-treatments are completed respectively, they are merged into the same downstream reaction chamber 12 for corresponding detection reactions, thereby further improving the application field of the biological detection chip and the adaptability to the working conditions, and making its operation efficiency better and better. Structural integration can be further improved.

It should be noted that, when only one side of the substrate 101 has a liquid channel, in order to ensure that the biochemical reactions in the reaction chamber 12 are independent of each other and do not interfere with each other, the volume of the diversion chamber 113 should be less than or equal to the reaction chamber 12, the precipitation chamber 124 and the buffer chamber The sum of the volumes of 121 is preferably larger than that of the reaction chamber 12; when there are liquid channels on both sides of the substrate 101, also in order to ensure the independence of the biochemical reactions in the reaction chamber 12, the two sides of the substrate 101 are divided into the guide chambers 113. The sum of the volumes should be less than or equal to the sum of the volumes of the reaction chamber 12 , the precipitation chamber 124 and the buffer chamber 121 , and preferably the volume is larger than that of the reaction chamber 12 .

The material of the substrate 101 can be one or a mixture of glass, silicon wafer, metal or polymer. The polymer can be PDMS (polydimethylsiloxa), PMMA (polymethylmethacrylate). Ester), PC engineering plastics, COC (copolymers of cycloolefin cycloolefin copolymer), PET (Polyethylene terephthalate), COP of Japan Zeon, ABS (Acrylonitrile butadiene Styrene copolymers acrylonitrile-butanediene) one or more of ethylene-styrene copolymers).

In addition, the outer wall of the base body 11 is covered with a sealing film 103 that is fitted with the feed opening 114 and the exhaust hole 13 . The sealing film 103 can reliably seal the feeding port 114 and the exhaust hole 13, so as to prevent the dust or impurities in the external environment from entering the inner chamber of the substrate 11 when the relevant biochemical reaction and detection are performed inside the biological detection chip, thereby Ensure the relative airtightness and sealing of each chamber, and ensure the accuracy and reliability of relevant test results.

It should be noted that, when it comes to practical application, considering the use and operation requirements under different working conditions, the above-mentioned sealing film 103 can be made of breathable materials or non-breathable materials, and the staff can flexibly choose according to the actual working conditions. , as long as it can meet the actual needs of the biological detection chip.

The working process of the biological detection chip provided by the present invention is as follows:

The relevant reagents and test samples required for test detection are added into the pretreatment chamber 111 through the sample inlet 114 , the air in the pretreatment chamber 111 is discharged to the external environment through the exhaust hole 13 , and then the sample inlet 114 is closed. Then put the chip into the auxiliary control device for heating and temperature control or light treatment. After the treatment is completed, use the centrifugal rotating device to drive the biodetection chip to rotate as a whole, so that the liquid in the pre-processing chamber 111 flows into the distribution chamber 112 under the action of centrifugal force, and Enter into each guide cavity 113 respectively, and at the same time, the gas in the distribution cavity 112 and the guide cavity 113 enters the pretreatment cavity 111 through the downstream conduit 116 to achieve pressure balance. When the liquid reaches the diversion cavity 113 at a low flow rate, although the diversion cavity 113 and the reaction cavity 12 are also in a connected state, because the pressure-limiting elbow 122 relies on its own conduction threshold to form a valve effect, at this time the liquid pressure It is less than the threshold value of the valve, therefore, the liquid can be prevented from entering the reaction chamber 12 through the pressure limiting elbow 122 in the non-test state. When the rotational speed of the centrifugal rotating device is increased and the liquid pressure in the diversion chamber 113 is greater than the conduction threshold of the pressure limiting elbow 122 , the liquid can enter the reaction chamber 12 for reaction, so as to complete the relevant biological detection operations.

In actual operation, the specific use method of the biological detection chip is as follows:

1) Add sample to the pre-processing chamber 111 of the biological detection chip through the sample adding port 114;

2) Seal the sample injection port 114 and the exhaust hole 13;

3) With the assistance of the supporting equipment, the sample is mixed and reacted with the preset reagents in the pretreatment chamber 111, and at the same time, the chip can be rotated by centrifugal rotating equipment such as a centrifuge, or the biological detection chip can be rotated by a temperature control device. The temperature of each internal chamber is controlled;

4) Low-speed centrifugation is performed on the biological detection chip by means of a centrifugal rotating device, and the liquid in step 3) is transferred to the distribution chamber 112, and with the assistance of the supporting equipment, the liquid is mixed and reacted with the preset reagents. This process Similarly, the rotation of the biological detection chip can be controlled as required, or the temperature in each chamber can be controlled;

5) using centrifugal rotating equipment to implement high-speed centrifugation on the biological detection chip, and further transfer the liquid in the distribution cavity 112 into each reaction cavity 12;

6) With the aid of the supporting equipment, the liquid is reacted with the preset reagent in the reaction chamber 12;

7) Detect and analyze the reaction results.

In order to further understand the technical content of this solution, the actual operation and use of the biological detection chip disclosed in the present invention will be further described in detail below by taking the detection of saliva as an example.

The saliva sample is added to the pretreatment chamber 111 of the integrated microfluidic chip through the sample injection port 114, and the virus lysis reagent is pre-embedded in the chamber; then the above-mentioned biological detection chip (hereinafter referred to as the chip) is passed through the supporting equipment at 37-95. ℃ (preferably 65 ℃) temperature environment heating for 1min～60min (preferably 30min) to obtain viral nucleic acid extract, use centrifugal rotating equipment to rotate the chip at 100rpm～3000rpm (preferably 1600rpm) and centrifuge for 10sec～60sec (preferably 45sec), the The above-mentioned viral nucleic acid extraction solution is transferred into the diversion chamber 113, and then through dissolution and diffusion, the preset constant temperature amplification reagent in the diversion chamber 113 is fully mixed with the viral nucleic acid extraction solution; Centrifuge the chip for 1 min, and evenly distribute the liquid in the diversion chamber 113 into each reaction chamber 12. The reaction chamber 12 is preset with primers that specifically react with the sample nucleic acid; The chip is heated for 30 to 60 minutes (preferably 60 minutes) in the environment, and a constant temperature amplification reaction is carried out in the reaction chamber 12. Finally, a matching instrument is used to detect the fluorescence in the reaction chamber 12 in real time and obtain the detection result.

It should be pointed out that the above-mentioned test equipment and environmental parameters are only to achieve the best test results under general experimental conditions, and each data parameter is only used for illustration purposes. In actual operation, different application conditions and specific tests are considered. For differences in detection requirements, those skilled in the art can flexibly adjust the specific values of each parameter according to the actual situation. In principle, as long as it can meet the specific needs of actual biological test detection.

The utility model has the following beneficial effects:

1) The biodetection chip has a pressure-limiting elbow that acts as a flow control valve, and the liquid can be transferred step by step only by controlling the centrifugal speed without the aid of an additional fluid control mechanism. The operation process is simple and easy, and no intermediate process is required. The manual operation intervention reduces the workload of the test personnel, and the test reactions can be carried out simultaneously in each reaction chamber, which realizes the multiple processing of related test operations and improves the detection efficiency;

2), the chip integration is high, the structure is simple, and the manufacturing and packaging costs are low;

3) The chip detection results are accurate and reliable, and the operation process is precise and controllable. It can be widely used in automatic nucleic acid extraction and multiple amplification detection of viruses or cells in various sample types such as saliva, throat swabs, and cervical swabs. It works normally in the ground environment and can also be applied to microgravity environments such as space stations, which greatly expands the scope of application of the product.

The biological detection chip provided by the present invention has been described in detail above. The principles and implementations of the present invention are described herein by using specific examples, and the descriptions of the above embodiments are only used to help understand the method and the core idea of the present invention. It should be pointed out that for those of ordinary skill in the art, without departing from the principles of the present utility model, some improvements and modifications can also be made to the present utility model, and these improvements and modifications also fall into the protection of the claims of the present utility model. within the range.

Claims (10)

1. A biological detection chip, characterized in that it comprises a base body (11), and the base body (11) is provided with a pre-processing cavity (111) which is sequentially communicated with the rotation center of the biological detection chip along the radial direction outward. ), a distribution cavity (112), a diversion cavity (113) and a reaction cavity (12), the diversion cavity (113) and the reaction cavity (12) are connected with a pressure-limited elbow (122), the Two inner sides of the upper inlet of the diversion cavity (113) are respectively designed with a closing structure (14) extending inwardly; It is also provided with an exhaust hole (13) which communicates the internal chambers and the external environment. 2 . The biological detection chip according to claim 1 , wherein the length of the closing structure ( 14 ) is less than half of the width of the guide cavity ( 113 ). 3 . 3. The biological detection chip according to claim 1, wherein the guide cavity (113) is along the circumferential direction of the centrifugal rotation circumference of the biological detection chip or along the centrifugal rotation circumference of the biological detection chip The extending directions of the involutes are arranged in sequence. 4 . The biological detection chip according to claim 1 , wherein the middle part of the pressure-limiting elbow (122) is a U-shaped pipe section, and two ends of the U-shaped pipe section are respectively connected with the guide cavity along the guide cavity. 5 . (113) The straight pipe section extending in the flow guiding direction, the main body straight pipe of the U-shaped pipe section is inclined relatively to the straight pipe section. 5 . The biological detection chip according to claim 4 , wherein a buffer chamber ( 121 ) is communicated between the diversion chamber ( 113 ) and the reaction chamber ( 12 ), and the pressure-limiting elbow ( 122) is communicated between the buffer cavity (121) and the diversion cavity (113), and a capillary (123) is communicated between the buffer cavity (121) and the reaction cavity (12). 6 . The biological detection chip according to claim 5 , wherein the distribution cavity ( 112 ) is communicated with a plurality of the guide cavities ( 113 ), and each of the guide cavities ( 113 ) passes through one The pressure limiting elbow (122) is communicated with one of the buffer chambers (121), and each of the buffer chambers (121) is communicated with one of the reaction chambers (12) through one of the capillary tubes (123). 7. The biological detection chip according to claim 1, wherein an upstream conduit (115) is communicated between the feed end of the distribution chamber (112) and the pretreatment chamber (111), and the The middle of the upstream conduit (115) has a large diameter buffer cavity (117). 8. The biological detection chip according to claim 7, characterized in that a downstream conduit ( 116), the exhaust hole (13) is arranged on the downstream conduit (116). 9. The biological detection chip according to claim 1, wherein a waste liquid chamber (118) is communicated with an end of the distribution chamber (112). 10 . The biological detection chip according to claim 1 , wherein a plurality of the pre-processing chambers ( 111 ) are provided on the substrate ( 11 ), and each of the pre-processing chambers ( 111 ) is formed by the The rotation centers of the biodetection chips are sequentially arranged and communicated radially outward.

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