WO2025023649A1 - Double-sided chip for nucleic acid extraction for pcr test, cartridge comprising same, and nucleic acid extraction device using same - Google Patents

Abstract

The present invention relates to a double-sided chip for nucleic acid extraction for a PCR test, a cartridge comprising same, and a nucleic acid extraction device using same. The double-sided chip for nucleic acid extraction for a PCR test includes: an input chamber into which a biological sample to be tested is inputted; an absorption chamber having, accommodated therein, an absorbent for absorbing and removing, from the sample, inhibitors which inhibit PCR performance; a nucleic acid chamber which accommodates therein a nucleic acid having passed through the absorption chamber so as to have the inhibitors removed therefrom; and a body unit which comprises the input chamber, the absorption chamber, and the nucleic acid chamber such that the sample sequentially passes through the input chamber, the absorption chamber, and the nucleic acid chamber.

Description

Double-sided chip for nucleic acid extraction for PCR test, cartridge containing same, and nucleic acid extraction device using same

The present invention relates to a double-sided chip for nucleic acid extraction for PCR testing, a cartridge containing the same, and a nucleic acid extraction device using the same. The present invention relates to a double-sided chip for nucleic acid extraction for PCR testing, a cartridge containing the same, and a nucleic acid extraction device using the same, which enables rapid and easy extraction of nucleic acids by moving a biological sample within a single chip, effectively removing foreign substances from the biological sample, and easily separating nucleic acids and buffers.

Polymerase Chain Reaction (PCR) is a method for amplifying specific target genetic material that is to be detected, and is a technology that amplifies a large amount of genetic material with the same base sequence from a small amount of genetic material. PCR is used to diagnose various types of genetic diseases by amplifying human nucleic acids, or to diagnose infectious diseases by applying it to nucleic acids of bacteria, viruses, and fungi.

In general, PCR is performed repeatedly through the following three steps. The three steps include: 1) a denaturing step in which a sample solution containing double-stranded DNA is heated to a specific temperature, for example, about 95°C, to separate the double-stranded DNA into single-stranded DNA; 2) an annealing step in which an oligonucleotide primer having a sequence complementary to a specific base sequence to be amplified is provided to the sample solution after the denaturing step, and the sample solution is cooled to a specific temperature, for example, 55°C, together with the separated single-stranded DNA, to bind the primer to the specific base sequence of the single-stranded DNA to form a partial DNA-primer complex; and 3) an extension (or amplification) step in which the sample solution is maintained at an active temperature of DNA polymerase, for example, 72°C, after the annealing step, to form double-stranded DNA based on the primer of the partial DNA-primer complex by DNA polymerase. By repeating the above three steps several times, a target nucleic acid having a specific base sequence can be exponentially amplified. However, in the case of RNA, a step of reverse transcribing RNA into DNA can be performed in advance in steps 1) to 3).

To perform this type of PCR, nucleic acids must be extracted from a biological sample.

As an example of a conventional nucleic acid extraction technique, there was a method of solubilizing a sample containing cells with SDS or proteinase K, and then denaturing and removing the proteins with phenol to purify the nucleic acids. However, the phenol extraction method requires many processing steps, so it takes a lot of time, and the nucleic acid extraction efficiency is greatly affected by the experience and skill of the researcher, so there was a problem that reliability was greatly reduced. Recently, kits using silica or glass fibers that specifically bind to nucleic acids have been used to solve this problem. Since the silica or glass fibers have a low binding ratio with proteins and cell metabolites, a relatively high concentration of nucleic acids can be obtained. This method has the advantage of being simple compared to the phenol method, but since it uses chaotropic reagents or ethanol that strongly inhibit enzymatic reactions such as polymerase chain reaction (PCR), these substances must be completely removed, and for this reason, the operation is very cumbersome and takes a long time.

In addition, in Korean Patent Registration No. 10-0454869, DNA extraction using a cell lysis buffer was performed as follows. 1) After culturing animal cells, the turbid solution containing the cells was centrifuged to recover the cells. 2) 300 ㎕ of cell lysis buffer was added to the recovered cells. 3) After adding the cell lysis buffer, it was left at 70℃ for 5 minutes. 4) In order to untangle denatured proteins, etc. in the lysis step so that they can pass through the filter easily, the lysed cells were pipetted 2-3 times. 5) After transferring the lysed cells to a filter made of silica membrane, centrifugation was performed at 13,000 rpm for 1 minute, the solution was discarded, and centrifugation was performed again. 6) 500 ㎕ of washing buffer was added to the filter and centrifugation was performed. To increase efficiency, the solution was discarded and centrifuged once more to completely remove the washing buffer. 7) 200 ㎕ of elution buffer was added to the filter and centrifuged, and the eluted solution was centrifuged once more in the filter to obtain a larger amount of genomic DNA. 8) The extracted DNA was observed after electrophoresis on a 12% agarose gel and stained with EtBr.

Thus, the conventional nucleic acid extraction method consists of the steps of 1) adding a cell lysis buffer to the cells to lyse the cells; 2) transferring the lysed cells of step 1 to a filter to fix the nucleic acids; 3) washing the filter of step 2; and 4) recovering the nucleic acids from the filter, and thus has the advantage of being able to extract nucleic acids with high reproducibility in a short period of time and steps. However, a centrifuge is used when transferring the lysed cells to the filter, washing the filter, or recovering the nucleic acids from the filter. The use of such a centrifuge has the effect of shortening the time, but has the problem of low portability and mobility and complicating the nucleic acid extraction process in the field.

In addition, there are nucleic acid extraction methods using magnetic beads, nucleic acid extraction methods using syringes and filters, nucleic acid extraction methods using direct lysis buffer (DLB), and nucleic acid extraction methods using Trizol, but the nucleic acid extraction method using magnetic beads requires a magnet to fix the nucleic acid to the wall of the tube, and the use of a pump or valve (automated equipment) or multiple tips and pipettes (manual) to remove the solution. In addition, the nucleic acid extraction method using a syringe and filter has a problem that the filter is damaged if a certain amount of force is applied during the process of transferring the nucleic acid using the syringe, making it difficult to extract the nucleic acid. The nucleic acid extraction method using direct lysis buffer (DLB) has a problem that the sensitivity greatly decreases due to dilution because the DLB (Direct lysis buffer) itself may contain PCR inhibitors, so it must be diluted by 1/10. In addition, the nucleic acid extraction method using Trizol has a problem that it uses harmful organic solvents such as phenol or chloroform.

Accordingly, there is a need for a device that can quickly and easily isolate nucleic acids from biological samples without using additional equipment.

The present invention aims to provide a double-sided chip for nucleic acid extraction for PCR testing, which effectively removes foreign substances from a biological sample while moving the biological sample within a single chip without using additional equipment, and easily separates nucleic acids and buffers, thereby quickly and easily extracting nucleic acids, a cartridge including the chip, and a nucleic acid extraction device using the chip.

A double-sided chip for nucleic acid extraction for PCR testing according to one aspect of the present invention comprises: an input chamber into which a biological sample to be tested is input; an absorption chamber containing an absorbent that absorbs and removes an inhibitor that inhibits PCR performance from the sample; a nucleic acid chamber containing nucleic acids from which inhibitors have been removed by passing through the absorption chamber; and a body part including the input chamber, the absorption chamber, and the nucleic acid chamber so that the sample sequentially passes through the input chamber, the absorption chamber, and the nucleic acid chamber.

In addition, it is preferable that the body portion is formed in a plate shape having a predetermined thickness and includes one side and an opposite side, and the absorption chamber is formed at a predetermined depth on either the one side or the opposite side.

In addition, it is preferable that the injection chamber has an injection port provided at the top through which the sample is injected, an outlet provided at the bottom through which the sample is discharged, and includes an inclined guide part that guides the sample to gather toward the outlet.

In addition, it is preferable that an air injection hole is formed in the injection chamber through which air is injected by an air pump, and that the sample is pushed and moved toward the absorption chamber by the air pressure provided through the air injection hole.

In addition, the absorption chamber includes a first absorption chamber, a second absorption chamber, and a third absorption chamber that are connected in the vertical direction, and the first, second, and third absorption chambers include a resin to which the inhibitor is adsorbed, and a mesh-structured barrier film is provided between the first absorption chamber and the second absorption chamber, and between the second absorption chamber and the third absorption chamber, so that the sample passes through the first, second, and third absorption chambers sequentially, and the resin is preferably prevented from moving by the barrier film.

In addition, it is preferable that the first absorption chamber, the second absorption chamber, or the third absorption chamber be filled with any one selected from among an anion exchange resin having a cathode, a chelating resin, or a cation exchange resin having a positive electrode that adsorbs the inhibitor.

In addition, it is preferable that a washing liquid storage chamber be provided between the injection chamber and the absorption chamber to accommodate a washing liquid for washing the absorbent filled in the absorption chamber.

In addition, the washing liquid storage chamber is filled with the washing liquid and includes a plurality of washing liquid flow paths arranged in parallel in one direction so that the washing liquid flows, and it is preferable that the washing liquid flows in a zigzag pattern along the washing liquid flow paths.

In addition, it is preferable that the washing liquid flow path includes: a first storage unit configured with a plurality of first channels having a first length and storing the washing liquid; a second storage unit configured with a plurality of second channels shorter than the first length and storing the washing liquid, and provided below the first storage unit; and a third storage unit configured with a plurality of second channels shorter than the second length and storing the washing liquid, and provided below the second storage unit.

In addition, it is preferable that the washing liquid storage chamber and the absorption chamber are formed on the same surface of the plate-shaped body portion.

In addition, the rear end of the absorption chamber includes a waste collection and nucleic acid separation chamber in which the washing liquid is collected and the nucleic acid is discharged; and it is preferable that the nucleic acid separated through the waste collection and nucleic acid separation chamber is collected in the nucleic acid chamber.

In addition, a washing liquid storage chamber is provided between the injection chamber and the absorption chamber, which accommodates a washing liquid for washing the absorbent filled in the absorption chamber; and a waste collection and nucleic acid separation chamber is provided at the rear end of the absorption chamber, which collects the washing liquid and discharges the nucleic acid; and it is preferable that at least one of the washing liquid storage chamber, the absorption chamber, and the waste collection and nucleic acid separation chamber is formed on at least one of one surface of the body portion having a predetermined thickness and the other surface opposite thereto.

In addition, it is preferable that the washing liquid storage chamber and the absorption chamber are formed on one surface of the body portion, and the waste collection and nucleic acid separation chamber are formed on the other surface of the body portion.

In addition, it is preferable that the body part further includes a tubular path forming a path through which a sample is transferred from the injection chamber to the nucleic acid chamber, and a valve for opening and closing the tubular path is provided.

In addition, the tubular path includes a first tube path provided between the injection chamber and the washing liquid storage chamber; a second tube path provided between the washing liquid storage chamber and the absorption chamber; and a third tube path provided between the absorption chamber and the sink waste collection and nucleic acid separation chamber; and it is preferable that the valve simultaneously opens or closes the first, second, and third tube paths.

Meanwhile, a double-sided chip for nucleic acid extraction for PCR testing according to another aspect of the present invention comprises: a plate-shaped body having a predetermined thickness; an absorption chamber formed on one surface of the body or the other surface opposite the one surface, and containing an absorbent that absorbs and removes an inhibitor that inhibits PCR performance from the sample; a waste collection and nucleic acid separation chamber formed on one surface of the body or the other surface opposite the one surface, and capturing a washing liquid that washes the absorption chamber, and passing through the absorption chamber to separate and discharge nucleic acids from which inhibitors have been removed.

Meanwhile, a cartridge including a double-sided chip according to another aspect of the present invention comprises a plate-shaped body part including an absorption chamber for absorbing and removing an inhibitor that inhibits PCR performance from a sample; a front cover coupled to one side of the body part; and a rear cover coupled to the front cover with the body part interposed therebetween; characterized in that nucleic acid is separated from the sample as the sample passes through the absorption chamber.

Meanwhile, a nucleic acid extraction device using a cartridge according to another aspect of the present invention comprises a plate-shaped body part including an absorption chamber for absorbing and removing an inhibitor that inhibits PCR performance from a sample; a front cover coupled to one side of the body part; and a rear cover coupled to the front cover with the body part interposed therebetween; a cartridge for separating nucleic acids from a sample as the sample passes through the absorption chamber; and

It is characterized by including an insertion part into which the cartridge is inserted and mounted, and a mounting part including a door provided on the insertion part for opening and closing the insertion part.

A double-sided chip for nucleic acid extraction for PCR testing according to an embodiment of the present invention, a cartridge including the same, and a nucleic acid extraction device using the same provide the effect of effectively removing foreign substances from a biological sample, easily separating nucleic acids and buffers, and quickly and easily extracting nucleic acids while moving the biological sample within a single chip without using additional equipment.

In addition, the present invention simplifies the nucleic acid separation procedure by easily separating nucleic acids on a single chip without using additional equipment such as a centrifuge for nucleic acid separation, and improves user convenience by making it easy to carry and move to the field.

Figure 1 is a perspective view of a double-sided chip for nucleic acid extraction according to one embodiment of the present invention.

Figure 2 is a cross-sectional view of the main part of Figure 1.

Figure 3 is a perspective view of Figure 1 from a different angle.

Figure 4 is a perspective view of the opposite side of Figure 1.

Figure 5 is a drawing showing the path along which the sample flows sequentially in the double-sided chip, indicated by numbers.

Figure 6 is a drawing showing the chambers through which the sample passes in sequence.

Figure 7 is a plan view of Figure 4.

Figure 8 is a plan view of Figure 3.

Figure 9 is a perspective view of Figure 1 from a different angle.

Figure 10 is a drawing showing the nucleic acid extraction in a nucleic acid chamber.

Figure 11 is a drawing showing a cartridge including a double-sided chip;

Figure 12 is an exploded perspective view of Figure 11.

Figure 13 is a drawing showing a state in which a double-sided chip is attached to a rear cover.

Figure 14 is a drawing showing a state where the valve closes the tubular path.

Figure 15 is a drawing showing a state where the valve has opened the tubular path.

Figure 16 is a drawing showing the valve rising by a pressure transmitting means.

Figure 17 is a drawing showing the cap sliding to open and close the injection chamber and the bottom surface of the cap.

Figure 18 is a perspective view of the cap.

Fig. 19 is a front view of Fig. 19.

Figure 20 is a drawing showing a cartridge mounted on a nucleic acid extraction device.

Figure 21 is a drawing showing how a pump connection part is connected to a cartridge.

Hereinafter, various embodiments of the present invention will be described in connection with the accompanying drawings. Various embodiments of the present invention may have various modifications and various embodiments, and thus specific embodiments are illustrated in the drawings and detailed descriptions related thereto are described. However, this is not intended to limit various embodiments of the present invention to specific embodiments, but should be understood to include all modifications and/or equivalents or substitutes included in the spirit and technical scope of various embodiments of the present invention. In connection with the description of the drawings, similar reference numerals have been used for similar components.

Expressions such as "includes" or "may include", which may be used in various embodiments of the present invention, indicate the presence of the disclosed corresponding function, operation or component, etc., and do not limit one or more additional functions, operations or components, etc. In addition, in various embodiments of the present invention, it should be understood that terms such as "includes" or "has" are intended to specify the presence of a feature, number, step, operation, component, part or a combination thereof described in the specification, but do not exclude in advance the possibility of the presence or addition of one or more other features, numbers, steps, operations, components, parts or combinations thereof.

When it is said that a component is "connected" to another component, it should be understood that while said component may be directly connected to said other component, there may also be other new components between said component and said other component. Conversely, when it is said that a component is "directly connected" or "directly connected" to another component, it should be understood that no other new components exist between said component and said other component.

The terms used in the various embodiments of the present invention are only used to describe specific embodiments and are not intended to limit the various embodiments of the present invention. The singular expression includes the plural expression unless the context clearly indicates otherwise.

Unless otherwise defined, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by one of ordinary skill in the art to which various embodiments of the present invention belong.

Terms defined in commonly used dictionaries should be interpreted as having a meaning consistent with their meaning in the context of the relevant technology, and will not be interpreted in an idealized or overly formal sense unless explicitly defined in various embodiments of the present invention.

The present invention relates to a double-sided chip for extracting nucleic acids for PCR tests, and relates to a chip and device for removing an inhibitor that interferes with PCR from a biological sample and separating and obtaining nucleic acids in order to perform PCR on the sample. Of course, the double-sided chip for extracting nucleic acids according to the present invention can be utilized not only in cases where extraction of nucleic acids is required for the purpose of diagnosing, treating, or preventing diseases, but also in various fields such as new drug development and detection of environmental hormones where extraction of nucleic acids from samples is required.

First, the term 'PCR (polymerase chain reaction) or polymerase chain reaction' used in this specification means a reaction that amplifies a specific target nucleic acid molecule using a heat-stable DNA polymerase. In addition to DNA polymerase, PCR may use a reaction mixture containing a primer (forward primer, reverse primer), which is an oligonucleotide that can specifically hybridize to a target nucleic acid, a deoxynucleotide triphosphate mixture (dNTP mixture), and a divalent ion such as Mg2+.

A 'primer' is used to initiate a PCR reaction, and refers to an oligonucleotide or polynucleotide that hybridizes complementarily to template DNA. A primer for a PCR reaction may be a pair of a forward primer (or sense primer) selected from a sense strand identical to the direction of progression of the genetic code of a nucleic acid molecule to be amplified, and a reverse primer (or antisense primer) selected from an antisense strand complementary to the sense strand.

The 'sample' refers to genetic material such as nucleic acid that is the target to be amplified or a biological solution containing such genetic material. In addition, the 'reaction reagent' may include fluorescent dyes, primers, etc. for detecting the target genetic material. The primer may be composed of a pair of primers of 15 to 30 bp in length that can bind to both ends of a specific region of the target gene. In addition, the DNA polymerase uses an enzyme that does not lose activity even at high temperatures of 90°C or higher.

Hereinafter, preferred embodiments of the present invention will be described in detail with reference to the attached drawings.

According to one embodiment of the present invention, a double-sided chip (1000) for nucleic acid extraction is provided, which forms a path and a chamber on both sides of a body part (10) through which a biological sample flows, so that an inhibitory substance (hereinafter, referred to as an inhibitor) that inhibits PCR is adsorbed and removed while passing through the path and chamber, and only nucleic acids are collected and separated into a nucleic acid chamber (300). According to this embodiment, the body part (10) is formed in a plate shape having a predetermined thickness, and an injection chamber (100), an absorption chamber (200), a nucleic acid chamber (300), a washing solution storage chamber (400), and a weight collection and nucleic acid separation chamber (500) to be described later are provided in the body part (10).

1. Injection chamber (100)

A biological sample to be tested is introduced into the injection chamber (100). Before dispensing the sample into the injection chamber, the sample is pretreated so that nucleic acids can be separated. The pretreatment is performed by injecting a predetermined dissolution buffer, for example, a lysis buffer, into the biological sample to destroy the cell wall and cause the nucleic acids to leak. The lysis buffer is a dissolution buffer used to destroy the cell wall. The sample to which the nucleic acids have been exposed is introduced into the injection chamber (100).

As shown in FIGS. 1 and 2, the injection chamber (100) is formed on one side of the body part (10) and forms a space in the vertical direction, and is formed so that the width becomes narrower as it goes downward. An injection port (101) into which the sample is injected is provided on the upper side of the injection chamber (100), and an outlet port (102) through which the sample is discharged is provided on the lower side. According to the present embodiment, an inclined guide portion (103) is formed to guide the sample to gather toward the outlet port (102).

According to the present embodiment, the sample injected into the injection chamber (100) moves to the washing liquid storage chamber (400) through the discharge port (102) formed at the bottom of the injection chamber (100) by air pressure injected from the top. The air is provided from a pump connected to the bottom of the body part (10), and the air provided by the pump is supplied to the upper side wall of the injection chamber (100). An air injection hole (104) into which air is injected by the pump is formed in the injection chamber (100). According to the present embodiment, the sample can be pushed and moved toward the absorption chamber (200) by the air pressure provided to the air injection hole (104).

2. Absorption chamber (200)

The absorption chamber (200) is provided to absorb and remove inhibitors that inhibit PCR from the sample. The inhibitors include salts, proteins, and other intracellular chemicals contained in the sample, and act as factors that interfere with the amplification of nucleic acids by the substances when performing PCR, and are removed by the absorbent contained in the absorption chamber (200). Since the inhibitors contained in the sample are absorbed and removed by the absorbent, and only the nucleic acids can be discharged and collected downward, the nucleic acids can be separated simply and quickly. The present invention provides a new concept of a separation device that separates nucleic acids without going through the process of fixing nucleic acids using a predetermined adsorption filter or the like to separate nucleic acids from a conventional sample and then re-separating the nucleic acids from the adsorption filter.

According to the present embodiment, the absorption chamber (200) includes first, second, and third absorption chambers (210, 220, and 230). The first, second, and third absorption chambers (210, 220, and 230) are filled with spherical resin particles for absorbing an inhibitor. The spherical resin particles absorb the inhibitor. According to the present embodiment, the first, second, and third absorption chambers (210, 220, and 230) are arranged in a vertical direction, and the sample passes through the first, second, and third absorption chambers (210, 220, and 230) sequentially.

According to the present embodiment, a mesh-structured barrier is formed at each boundary of the first, second, and third absorption chambers (210, 220, 230) to prevent the resin contained in one chamber from moving to another chamber. According to the present embodiment, a fastening groove (203) into which the barrier is inserted is formed between the first absorption chamber (210) and the second absorption chamber (220) and between the second absorption chamber (220) and the third absorption chamber (230). In addition, according to the present embodiment, a fastening groove (203) is also formed between the absorption chamber inlet (201) and the first absorption chamber (210) and between the third absorption chamber (230) and the absorption chamber outlet (202), so that a barrier is installed to prevent the resin from leaking.

The above first, second, and third absorption chambers (210, 220, and 230) may be selected to include any one of a cathodic anion exchange resin, a positive cation exchange resin, and a chelate resin. The chelate resin has a weak anode and has a property of adsorbing (capturing) metal divalent ions. In addition, the anion exchange resin, chelate resin, and cation exchange resin are formed by including a spherical resin, and the inhibitor is adsorbed onto the spherical resin. The diameter of the spherical resin may be formed to be approximately 0.18 mm to 0.3 mm.

The above absorption chamber (200) has three chambers, each of which has a polarity, so that an inhibitor having a polarity in the biological sample can be absorbed and removed in the first, second, and third absorption chambers (210, 220, and 230). According to the present embodiment, the first absorption chamber (210) includes an anion exchange resin, the second absorption chamber (220) includes a chelate resin, and the third absorption chamber (230) includes a cation exchange resin. As described above, the first, second, and third absorption chambers (210, 220, and 230) are arranged sequentially from top to bottom, so that the sample passes through the anion exchange resin, the chelate resin, and the cation exchange resin sequentially.

According to the present embodiment, the volumes of the anion exchange resin, the chelate resin, and the cation exchange resin can be prepared substantially as 1:1:1. Of course, the volume ratio of the anion exchange resin, the chelate resin, and the cation exchange resin is not limited to 1:1:1. The substances included in the biological sample are polar, so that they can be absorbed in each layer while passing through the first, second, and third absorption chambers (210, 220, and 230), and only the nucleic acid can flow downward. The anion exchange resin can include at least one of TMA (Trimethylamine), DMEA (Dimethylethanolamine), and Tertiary Amine, and the chelate resin can include at least one of a carboxyl group (-COOH), an iminodiacetate acid chelate, and the like. In addition, the cation exchange resin can include at least one of a sulfate group (-SO3H), a carboxyl group (-COOH), and a sulfonic acid group (-SO3H).

An absorption chamber outlet (202) is formed at the lower side of the third absorption chamber, and an inhibitor is removed from the sample through the absorption chamber outlet (202) and the remaining nucleic acid is discharged downward. According to the present embodiment, the sample moves inside the chip at a speed of 15 μl/sec or less, and as a result, the flow rate of the nucleic acid discharged through the absorption chamber outlet (202) becomes 15 μl per second or less. The flow rate per unit time of the nucleic acid discharged through the absorption chamber outlet (202) can be controlled by appropriately forming the cross-sectional area of the absorption chamber outlet (202) through which the nucleic acid passes. As the cross-sectional area of the above absorption chamber outlet (202) becomes relatively larger, the discharge amount of the nucleic acid may increase. However, since the speed at which the sample passes through the absorption chamber (200) relatively increases, it may not be possible to secure enough time for the inhibitor to be sufficiently adsorbed in the first, second, and third absorption chambers (210, 220, 230). Therefore, it was confirmed that the inhibitor could be sufficiently removed when the discharge flow rate of the nucleic acid was adjusted to 15 μl per second or less.

3. Nucleic acid chamber (300)

The nucleic acid chamber (300) receives nucleic acids from which inhibitors have been removed by passing through the absorption chamber (200). As illustrated in FIGS. 1 and 2, according to the present embodiment, the nucleic acid chamber (300) has a space whose width narrows from the top to the bottom. As illustrated in FIG. 10, a nucleic acid chamber inlet (310) through which nucleic acids are introduced is formed in the nucleic acid chamber (300). The nucleic acids collected in the nucleic acid chamber (300) can be extracted by a device such as a pipette (330) through an extraction port (320) formed opposite the nucleic acid chamber inlet (310).

According to the present embodiment, the sample passes through the injection chamber (100), the absorption chamber (200), and the nucleic acid chamber (300) sequentially, and the body part (10) is provided with the injection chamber (100), the absorption chamber (200), and the nucleic acid chamber (300), so that the process of separating and storing nucleic acids from the sample is possible in one body part (10).

4. Washing fluid storage chamber (400)

The washing liquid storage chamber (400) is a chamber provided between the injection chamber (100) and the absorption chamber (200), and accommodates a washing liquid for washing an absorbent filled in the absorption chamber (200). The absorption chamber (200) is filled with a polar resin, and each chamber is filled with a preservation buffer to maintain the polarity of the first, second, and third absorption chambers (210, 220, 230) forming the absorption chamber (200). The washing liquid is provided to wash the preservation buffer.

According to the present embodiment, the washing liquid storage chamber (400) is formed on one side of the body part (10) and is formed on the same side as the side on which the absorption chamber (200) is formed. When the sample injected into the injection chamber (100) is pushed out by air pressure, the washing liquid stored in the washing liquid storage chamber (400) moves toward the absorption chamber (200) by the movement pressure of the sample and air, and the absorbent of the absorption chamber (200) is washed first.

As illustrated in FIG. 4, the washing liquid storage chamber (400) includes a plurality of washing liquid flow paths (440) arranged in parallel in one direction so that the washing liquid can flow. As illustrated in FIG. 7, the washing liquid flows into the washing liquid storage chamber inlet (450) on the upper side, passes through the washing liquid flow path (440), and then moves to the absorption chamber (200) through the washing liquid storage chamber outlet (460). The washing liquid flows in a zigzag pattern from upper to lower along the washing liquid flow path (440). According to the present embodiment, the washing liquid flow path (440) includes a first path (401), a second path (402), and a third path (403) having different lengths.

According to the present embodiment, the washing liquid storage chamber (440) includes a first storage section (410) formed of a plurality of first channels (401) having a first length to store the washing liquid, a second storage section (420) formed of a plurality of second channels (402) shorter than the first length to store the washing liquid and provided below the first storage section (410), and a third storage section (430) formed of a plurality of second channels (403) shorter than the second length to store the washing liquid and provided below the second storage section (430). According to the present embodiment, the first, second, and third storage sections (410, 420, 430) are sequentially provided from the top to the bottom. The above first, second, and third storage units (410, 420, 430) are formed with a plurality of horizontal flow paths (401, 402, 403) in a zigzag manner, so that sufficient washing liquid can be stored.

5. Waste collection and nucleic acid separation chamber (500)

The waste collection and nucleic acid separation chamber (500) is provided to collect the washing liquid discharged from the washing liquid storage chamber (400) at the rear end of the absorption chamber (200) and to separate and discharge the nucleic acid. The nucleic acid separated by the waste collection and nucleic acid separation chamber (500) is collected in the nucleic acid chamber (300). The waste collection and nucleic acid separation chamber (500) may be formed on one or the other surface of the body portion (10).

Therefore, according to the present invention, at least one of the absorption chamber (200), the washing liquid storage chamber (400), and the waste collection and nucleic acid separation chamber (500) may be formed on at least one of one surface of the body part (10) having a predetermined thickness and the other surface opposite thereto. That is, at least one of the absorption chamber (200), the washing liquid storage chamber (400), and the waste collection and nucleic acid separation chamber (500) is formed on one surface of the body part (10) having a predetermined thickness, and the remainder of the absorption chamber (200), the washing liquid storage chamber (400), and the waste collection and nucleic acid separation chamber (500) that are not formed on the one surface may be formed on the other surface opposite thereto. According to the present embodiment, the washing liquid storage chamber (400) and the absorption chamber (200) are formed on one surface of the body part (10), and the waste collection and nucleic acid separation chamber (500) is formed on the other surface of the body part (10).

However, the absorption chamber (200) may be formed on one side or the other side of the body part (10), and the waste collection and nucleic acid separation chamber (500) may also be formed on one side or the other side of the body part (10). That is, in one embodiment, the absorption chamber (200) and the waste collection and nucleic acid separation chamber (500) may be formed on the same side of the body part (10).

Referring to FIGS. 8 and 9, the waste collection and nucleic acid separation chamber (500) is provided to collect waste and separate nucleic acids by having the washing liquid passing through the absorption chamber (200) fill the waste chamber (510) and subsequently moving the nucleic acids flowing through the inflow path (570) through the nucleic acid separation path (530).

The above waste collection and nucleic acid separation chamber (500) includes an inlet passage (570), a washing liquid collection passage (520), a nucleic acid separation passage (530), and a waste chamber (510).

The above inflow path (570) extends from the upper side to the lower side and is a path through which washing liquid and nucleic acid flow. According to the present embodiment, the washing liquid first flows into the inflow path (570), and the nucleic acid flows in following the washing liquid. The inflow path (570) includes a curved path (560) having a predetermined curvature at its end.

The washing liquid collection channel (520) is connected to extend downward from the end of the inflow channel (570) so that the washing liquid can flow. According to the present embodiment, the washing liquid collection channel (520) is connected to a curved channel (560) provided at the end of the inflow channel (570) and extends downward.

The nucleic acid separation channel (530) is extended in a different direction from the washing solution collection channel (520) at the end of the inflow channel (570), thereby forming a channel through which the nucleic acid flows. According to the present embodiment, the nucleic acid separation channel (530) is extended upward in the opposite direction of the washing solution collection channel (520) while being connected to the curved channel (560). The nucleic acid separation channel (530) is provided with a resistance protrusion (590) protruding from the inner surface. The resistance protrusion (590) has the function of inducing the washing solution to move toward the washing solution collection channel (520) downward when the washing solution is initially introduced.

According to the present embodiment, a resistance portion (580) that generates resistance when the washing liquid flows is formed on the round inner surface where the inflow path (570) and the nucleic acid separation path (530) are connected to each other, that is, on the inner surface of the curved path (560). According to the present embodiment, the resistance portion (580) may be formed as a protrusion that protrudes like a sawtooth shape on the inner surface that is satisfied from the end of the curved path (560) toward the nucleic acid separation path (530). The protrusions may be formed at a predetermined interval. The washing liquid flows toward the nucleic acid separation path (530) that extends downward by gravity, and at this time, the washing liquid collides with the resistance portion (580) while passing through the curved path (560), so that the washing liquid can be more easily guided and flow toward the lower waste chamber (510).

In addition, according to the present embodiment, as illustrated in FIG. 8, the width (W2) of the washing solution collection channel (520) is formed wider than the width (W1) of the nucleic acid separation channel (530). The washing solution flows into the inflow channel (570), passes through the curved channel (560), and can flow more smoothly toward the wider washing solution collection channel (520) at the end of the curved channel (560). In addition, a washing solution induction channel (524) is formed in the washing solution collection channel (520) that has a narrower width than the width of the washing solution collection channel (520) and is sunken deeper than the bottom of the washing solution collection channel (520). The above washing liquid induction path (524) allows the washing liquid to easily flow downward from the end of the curved path (560) toward the waste chamber (510) by capillary phenomenon.

The above-described waste chamber (510) is provided at an end of the washing liquid collection path (520) to provide a space in which the washing liquid is filled. According to the present embodiment, the washing liquid flows in from the bottom of the waste chamber (510) and fills the space of the waste chamber (510). According to the present embodiment, as illustrated in FIG. 9, a support rib (501) that protrudes vertically from the bottom surface is formed on the inside of the waste chamber (510). According to the present embodiment, a plurality of the support ribs (501) are provided. According to the present embodiment, one side and the other side of the body part (10) are sealed by adhering a film. The support ribs (501) prevent the surface of the film from sinking into the space of the waste chamber (510) when sealing the other side of the body part (10) with the film. When the above film is adhered to the other surface of the body part (10), if the area of the space secured by sinking into the space of the waste chamber (510) is reduced, the storage space of the washing liquid is reduced accordingly, which may cause a problem in which the washing liquid is discharged together with the nucleic acid through the nucleic acid separation path (530). Therefore, the support rib (501) ensures that the space of the waste chamber (510) can secure a space in which the washing liquid can be stored.

According to the present embodiment, a pressure-providing portion (540) is provided on the upper side of the waste chamber (510) so that the washing liquid is filled and the nucleic acid is no longer introduced into the waste chamber (510) by the pressure of the washing liquid. The pressure-providing portion (540) includes a microchannel (550) formed on the upper side of the waste chamber (510). The width of the microchannel (550) is narrower than that of the inflow channel (570), the curved channel (560), the waste collection channel (520), and the nucleic acid separation channel (530). A plurality of micro-spaces (541) are included on the microchannel (550).

When the washing liquid is filled in the waste chamber (510), the internal pressure of the waste chamber (510) increases by the microchannel (550) and the microspace (541). Some of the washing liquid may penetrate into the microchannel (550) and the microspace (541). The microspace (541) is provided in case sufficient pressure is not applied to a portion of the microchannel (550) connected to the waste chamber (510), and even if the washing liquid penetrates through the microchannel (550) near the waste chamber (510), pressure is applied again by the remaining microchannel (550) and the microspace (541).

In addition, the microchannel (550) and microspace (541) prevent the pressure of the waste chamber (510) from increasing rapidly. When the pressure increases by the pressure providing unit (540), the washing liquid is no longer transported toward the waste chamber (510), so that the nucleic acid subsequently introduced through the inflow channel (570) can flow upward through the nucleic acid separation channel (530) due to the repulsive force. Through this process, the washing liquid that washes away the absorbent of the absorption chamber (200) is collected and separated in the waste chamber (510), and only the nucleic acid can move to the nucleic acid chamber (300) through the nucleic acid separation channel (530).

Meanwhile, according to another aspect of the present invention, the double-sided chip (1000) may be manufactured and provided in the form of a cartridge (2000).

As illustrated in FIGS. 11 and 12, a cartridge (2000) according to the present embodiment includes a body portion (10), a front cover (2100) and a rear cover (2200) which are respectively coupled to one side and the other side of the body portion (10). The body portion (100) is positioned between the front cover (2100) and the rear cover (2200), and separates nucleic acids from a sample as the sample passes through the absorption chamber (200).

The above body part (10) is formed in a plate shape with a predetermined thickness and includes an injection chamber (100), an absorption chamber (200), a nucleic acid chamber (300), a washing liquid storage chamber (400), and a waste collection and nucleic acid separation chamber (500). A repetitive description of each chamber formed in the body part (10) is omitted as described above.

The body part (10) further includes a tubular flow path (700) in addition to the flow path formed on the surface of the body part (10) so that a sample can be transferred from the injection chamber (100) to the nucleic acid chamber (300). According to the present embodiment, a valve (600) for opening and closing the tubular flow path (700) is provided. As illustrated in FIG. 12, the tubular flow path (700) is connected by having both ends inserted into the tube flow path connecting portion (13) provided in the body part (10).

The above-described tube-shaped path (700) includes a first tube path provided between the injection chamber (100) and the washing liquid storage chamber (400), a second tube path provided between the washing liquid storage chamber (400) and the absorption chamber (200), and a third tube path provided between the absorption chamber (200) and the waste waste collection and nucleic acid separation chamber (500). In addition, according to the present embodiment, the valve (600) simultaneously opens or closes the first, second, and third tube paths.

As shown in FIGS. 14 and 15, the valve (600) is coupled to the body part (10) so as to be movable in the vertical direction. As shown in FIG. 14(c), when the valve (600) is in the first position, the pressure rib (610) formed in the valve (600) presses the tubular flow path (700) to maintain the flow path in a closed state. That is, the tubular flow path (700) is in a state in which the valve (600) is pressed in the first position as shown in FIGS. 14(a) and 14(b). Then, as shown in FIG. 15, when the valve (600) moves to a second position higher than the first position, the pressure rib (610) pressing the tubular flow path (700) moves upward, opening the tubular flow path (700) so that the sample can flow in one direction.

A groove is formed in a rear cover (2200) coupled to one side of the body portion (10), and a valve exposure hole (2210) is formed on an upper side of the groove to expose the valve (600) so as to transmit a pressing force to move the valve upward. As illustrated in FIG. 16, when a cartridge (2000) is inserted into a nucleic acid extraction device (3000) and a nucleic acid extraction operation is performed, a pressing force transmission means (3600) provided in the nucleic acid extraction device (3000) transmits a force to move the valve (600) upward through the valve exposure hole (2210). As the valve (600) moves upward by the pressing force, the tubular flow path (700) is opened.

In addition, a cap (2300) for opening and closing the injection port (101) of the injection chamber (100) is coupled to the upper side of the body part (10). As illustrated in FIG. 17, the gap (2300) is slidably coupled to the upper surface of the body part (10). The cap (2300) includes a guide projection (2370) that is inserted into and guided by a sliding groove (14) formed in the body part (10), and a sealing member (2330) that seals the edge of the injection port (101) at a position where the injection port (101) is closed.

As shown in FIGS. 17 and 18, the cap (2300) is formed with a cut groove (2310) that faces each other and extends in the longitudinal direction. A catch (2320) is formed protrudingly in the cut groove (2310). A protrusion (15) is formed in the width direction on the upper surface of the body (10), and when the cap (2300) is slid to close the inlet (101), the cut groove (2310) is elastically deformed so that the catch (2320) can pass over the protrusion (15). When the cap (2320) moves to the upper side of the inlet (101), a sealing member (2330) such as an O-ring that is coupled to the lower surface of the cap (2300) is placed on the edge of the inlet (101) to seal the inlet (101), and the protrusion (15) is maintained in a state of being caught by the catch (2320) so that the cap (2300) closes the inlet (101).

As illustrated in FIG. 18, the cap (2300) includes a handle (2340), a guide piece (2360) on which the guide projection (2370) is formed, and a position control projection (2350) that regulates the sliding range of the cap (2300). As illustrated in FIG. 19, the guide piece (2360) extends downward from both ends of the cap (2300) and is coupled to the upper side of the body part (10), and the guide projection (2370) formed on the inner side of the guide piece (2360) is inserted into the sliding groove (14) of the body part (10). The position control projection (2350) is formed in the longitudinal direction on the upper surface of the cap (2300) so that its end can come into contact with the front cover (2100) or the rear cover (2200) to limit the movement range of the cap (2300).

Meanwhile, according to another aspect of the present invention, a nucleic acid extraction device (3000) that receives the cartridge and extracts nucleic acids is proposed. The extraction device (3000) includes a cartridge (2000) including the double-sided chip described above, and a mounting portion (3100) on which the cartridge (2000) is mounted. Since the configuration of the cartridge (2000) has been described above, a repetitive description will be omitted.

The above-mentioned mounting portion (3100) may include an insertion portion (3200) into which the cartridge (2000) is inserted and mounted, a door (3300) for opening and closing the insertion portion (3200), a select bar (3400) for controlling the nucleic acid extraction device (3000), and a display portion (3500) for externally displaying whether the nucleic acid extraction device (3000) is operating.

As shown in Fig. 20, according to the present embodiment, the insertion portion (3200) is provided so that two cartridges (2000) can be inserted. Of course, the number of cartridges (2000) that can be accommodated is not limited thereto.

The above select bar (3400) can be implemented to enable selection of the on/off operation and operation time of the nucleic acid extraction device (3000). The display unit (3500) is provided on the upper surface of the mounting unit (3100) to visually check the operation progress status of the nucleic acid extraction device (3000). According to the present embodiment, the select bar (3400) is provided in a knob type, and the nucleic acid extraction device (3000) programmed under predetermined conditions can be operated by rotating the select bar (3400).

As illustrated in FIG. 21, the mounting portion (3100) is provided with a pump connection portion (3700) for supplying air to the cartridge (2000). A docking portion (11) for receiving air pressure from an air pump is provided in the body portion (10) of the cartridge (2000), and a docking hole (2230) is provided to expose the docking portion (11) at the lower side when the front cover (2100) and the rear cover (2200) are coupled to each other. When the cartridge (200) is inserted into the insertion portion (3200) and mounted, as illustrated in FIG. 21, the upper portion of the pump connection portion (3700) is inserted into the docking portion (11), and a pump is connected to the lower portion.

Below, the function and effect of the double-sided chip for nucleic acid extraction for PCR testing according to the above configuration are explained based on experimental results.

The present invention relates to a double-sided chip (1000), a cartridge (2000), and a nucleic acid extraction device (3000) for extracting nucleic acids. The present invention improves the nucleic acid extraction process to be simpler by removing inhibitory substances using an absorbent (resin type) in an absorption chamber (200) rather than the conventional concept of capturing and concentrating nucleic acids using a column method or magnetic beads and then extracting nucleic acids again from the beads, thereby dramatically speeding up and simplifying the nucleic acid extraction process. In addition, the nucleic acids can be automatically separated from a buffer (washing solution) in a waste collection and nucleic acid separation chamber (500) by using the difference in pressure.

- Flow of sample in double-sided chips

After a sample is introduced into the injection chamber (100), the cartridge (2000) is mounted on the nucleic acid extraction device (3000) to perform the nucleic acid extraction process. Air is supplied into the interior of the injection chamber (100) through the docking portion (11) of the cartridge (2000) from a pump connected to the nucleic acid extraction device (3000).

As shown in FIGS. 5 and 6, air is supplied to 0, comes out to 1, and flows to 2. 2 is connected to 3. The air flowing to 2 moves to 3 on the opposite side. The air moves from 3 to 4, moves to the opposite side again through 5, which is connected to 4, and rises to 6, and then is supplied to the injection chamber (100). The sample received in the injection chamber (100) is discharged to 7 by air pressure. The sample moves to 8, comes out to 9 on the opposite side, and moves from 9 to 10 along the tube-shaped path. The sample introduced to 10 comes out to 11 on the opposite side, flows into the washing liquid storage chamber (400), and then flows. At this time, the washing liquid stored in the washing liquid storage chamber (400) moves to No. 12 due to the movement pressure of the sample and is discharged to No. 13 on the opposite side. Nos. 13 and 14 are connected by a tube-shaped path. The washing liquid passing through No. 14 flows again into No. 15 on the opposite side and washes the resin contained in the first, second, and third absorption chambers (210, 220, 230) while passing through the absorption chamber (200). The washing liquid passing through the absorption chamber (200) is discharged to No. 16 and passes through Nos. 17 and 18. Then, the washing liquid passes through Nos. 19 and 20 and flows into No. 21. At this time, Nos. 17 and 18 are connected by a tube-shaped path.

The washing liquid introduced into No. 21 flows downward and enters the waste collection and nucleic acid separation chamber (500). As described above, the washing liquid first flows downward through the washing liquid collection channel (520) to fill the waste chamber (510), and when the waste chamber (510) is filled with the washing liquid, the nucleic acid (nucleic acid to which the inhibitor is adsorbed in the absorption chamber (200) and which is discharged from the absorption chamber (200)) flowing following the washing liquid flows toward No. 23 as pressure is applied by the narrow microchannel (550) of No. 22. The nucleic acid discharged through No. 23 exits through No. 24 on the opposite side and flows into the nucleic acid chamber (300) through No. 25 to be extracted. The user can capture the nucleic acid extracted through No. 26 using a pipette (330), etc.

- Experimental verification of the function and effectiveness of the absorption chamber

When the sample does not contain an inhibitor, and the nucleic acid is passed through a polar resin layer and then PCR is performed, it was confirmed that the effect on the number of PCR cycles for detecting a specific base sequence is minimal when performing PCR using Direct Lysis Buffer (DLB) prepared using a conventional lysis buffer and when performing PCR after passing through at least one absorbent (an absorption chamber including at least one of the first, second, and third chambers can be configured). The above PCR cycle is defined as the following three steps as one cycle. The above three steps mean: 1) a denaturing step of heating a sample solution containing double-stranded DNA to a specific temperature, for example, about 95°C, to separate the double-stranded DNA into single-stranded DNA; 2) an annealing step of providing an oligonucleotide primer having a sequence complementary to a specific base sequence to be amplified in the sample solution after the denaturing step, and cooling the solution together with the separated single-stranded DNA to a specific temperature, for example, 55°C, to bind the primer to the specific base sequence of the single-stranded DNA to form a partial DNA-primer complex; and 3) an extension (or amplification) step of forming double-stranded DNA based on the primer of the partial DNA-primer complex by maintaining the sample solution at an active temperature of DNA polymerase, for example, 72°C, after the annealing step, by DNA polymerase. PCR exponentially amplifies target nucleic acids having a specific base sequence by repeating the above three steps several times to the extent that the detection of a specific base sequence is possible. In Table 1 below, the CT value of 29.56 means that the average number of repetitions is 29.56. From these experimental results, it was confirmed that although the nucleic acid has a weak cathode, when PCR was performed by passing it through an anion exchange resin, a chelating resin, and a cation exchange resin respectively, and when these absorbents were configured in three stages and passed through a three-stage chamber, the number of repetitions of the PCR cycle that enables the detection of a specific gene was only slightly different from the prior art, and thus the nucleic acid was not affected by the resin layer.

CT values of clinical specimens without inhibitors DLB 29.56 Cation exchange resin 29.84 Kelate resin 29.77 Anion exchange resin 30.15 3-stage resin of anion, chelate, and cation 30.63

And, when the sample contains an inhibitor, as shown in Table 2 below, the sample prepared using the conventional dissolution buffer contained an inhibitor, so even if the above three steps for PCR were performed, the amplification of a specific base sequence was not achieved due to the inhibitor. However, as shown in Table 2, when the sample was passed through an anion exchange resin, a chelating resin, and a cation exchange resin and the three steps for PCR were repeated, it was confirmed that a specific base sequence was amplified to a detectable level. In particular, when the absorption chamber was configured as a three-stage chamber with resin layers including an anion exchange resin, a chelating resin, and a cation exchange resin from the upper side to the lower side, it was confirmed that the number of PCR cycle repetitions for detecting a specific base sequence was the shortest. That is, when the absorption chamber is configured to sequentially pass through the three-stage resin layers, the amplification of a specific base sequence can be completed quickly.

CT values of clinical specimens with inhibitors DLB(control) 0 Cation exchange resin layer (123) 36.68 Kelate resin layer 41.34 Anion exchange resin layer (121) 39.56 Three-layer resin layer of anion, chelate, and cation 35.13

According to the present embodiment, the absorption chamber (200) is sequentially arranged with first, second, and third absorption chambers (210, 220, 230) including a cathode-bearing anion exchange resin, a chelate resin, and a cathode-bearing cation exchange resin. In another embodiment, the absorption chamber (200) may be formed by selecting at least one of the first, second, and third absorption chambers (210, 220, 230). That is, the absorption chamber may be formed by a combination of at least one, two, or three layers.

According to an embodiment of the present invention, the volume ratio of the anion exchange resin, the chelate resin, and the cation exchange resin formed of spherical resin particles included in the first, second, and third absorption chambers (210, 220, and 230) is substantially 1:1:1. In addition, the flow rate of the sample penetrating the inside of the absorption chamber is adjusted to 15 μl/sec or less. Since the flow rate of the sample penetrating the inside of the absorption chamber is adjusted to 15 μl/sec or less, the flow rate of nucleic acid discharged to the lower part of the absorption chamber is adjusted to 15 μl per second or less, and the inhibitor can be sufficiently absorbed into the absorbent of the spherical particles while the sample passes through the first, second, and third absorption chambers (210, 220, and 230). When the flow rate of the nucleic acid discharged exceeds 15 μl per second, the movement speed of the sample becomes fast, so that the inhibitor cannot be sufficiently absorbed.

According to an embodiment of the present invention, the amount of nucleic acid collected in the nucleic acid chamber (300) can be secured substantially the same as the amount of nucleic acid injected into the injection chamber (100). For example, when 500 μl of the biological sample is injected, the inhibitor contained in the sample is removed and the flow rate substantially discharged to the lower part of the absorption chamber (200) is maintained at approximately 500 μl, and the inside is obtained in a state where the inhibitor is removed and only the nucleic acid is contained. This significantly improves the secured flow rate of the sample containing only the nucleic acid compared to the prior art.

In this way, the double-sided chip for nucleic acid extraction for PCR testing, the cartridge including the same, and the nucleic acid extraction device according to the embodiment of the present invention effectively remove foreign substances from the biological sample while moving the biological sample within a single chip without using additional equipment, and easily separate the nucleic acid and buffer, thereby providing the effect of quickly and easily extracting the nucleic acid. In addition, the present invention excludes the use of additional equipment such as a centrifuge for nucleic acid separation, so that it is easy to carry and move to the site, and can improve the convenience of the user.

Above, the present invention has been described in detail with reference to preferred embodiments, but the present invention is not limited to the above embodiments, and various modifications can be provided within a scope that does not depart from the scope of the present invention.

Claims (15)

An injection chamber into which the biological sample to be tested is injected; An absorption chamber containing an absorbent that absorbs and removes an inhibitor that inhibits PCR performance from the sample; A nucleic acid chamber in which nucleic acid from which the inhibitor has been removed is received by passing through the absorption chamber; and A double-sided chip for nucleic acid extraction for PCR testing, comprising a body part including the injection chamber, the absorption chamber, and the nucleic acid chamber, such that the sample passes through the injection chamber, the absorption chamber, and the nucleic acid chamber sequentially. In the first paragraph, A double-sided chip for nucleic acid extraction for PCR testing, characterized in that the body portion is formed in a plate shape having a predetermined thickness and includes one side and an opposite side, and the absorption chamber is formed at a predetermined depth on either the one side or the opposite side. In the first paragraph, The above injection chamber is, An inlet is provided on the upper side into which the sample is injected, A discharge port is provided on the lower side through which the sample is discharged. A double-sided chip for nucleic acid extraction for PCR testing, characterized by including an inclined guide section that guides the sample to gather toward the discharge port. In the first paragraph, A double-sided chip for nucleic acid extraction for PCR testing, characterized in that an air inlet hole is formed in the injection chamber into which air is injected by an air pump, and the sample is pushed and moved toward the absorption chamber by the air pressure provided through the air inlet hole. In the first paragraph, The above absorption chamber includes a first absorption chamber, a second absorption chamber, and a third absorption chamber connected in a vertical direction, The above first, second and third absorption chambers contain a resin to which the inhibitor is adsorbed. A double-sided chip for nucleic acid extraction for PCR testing, characterized in that a mesh-structured barrier is provided between the first absorption chamber and the second absorption chamber, and between the second absorption chamber and the third absorption chamber, so that the sample passes through the first, second, and third absorption chambers sequentially, and the resin is prevented from moving by the barrier. In paragraph 5, A double-sided chip for nucleic acid extraction for PCR testing, characterized in that the first absorption chamber, the second absorption chamber, or the third absorption chamber is filled with one selected from among an anion exchange resin, a chelating resin, or a cation exchange resin having a negative electrode that adsorbs the inhibitor. In the first paragraph, A double-sided chip for nucleic acid extraction for PCR testing, characterized in that a washing solution storage chamber is provided between the injection chamber and the absorption chamber to accommodate a washing solution for washing the absorbent filled in the absorption chamber. In Article 7, The above washing liquid storage chamber is filled with the washing liquid and includes a plurality of washing liquid flow paths arranged in parallel in one direction to allow the washing liquid to flow. A double-sided chip for nucleic acid extraction for PCR testing, characterized in that the washing solution flows in a zigzag pattern along the washing solution flow path. In Article 8, The above washing liquid storage chamber is, A first storage section comprising a plurality of first euros having a first length and storing the washing liquid: A second storage section, which is formed of a plurality of second euros shorter than the first length and stores the washing liquid, and is provided below the first storage section; A double-sided chip for nucleic acid extraction for PCR testing, characterized in that it comprises a third storage section provided below the second storage section, the third storage section comprising a plurality of second euros shorter than the second length and storing the washing liquid. In Article 7, A double-sided chip for nucleic acid extraction for PCR testing, characterized in that the washing liquid storage chamber and the absorption chamber are formed on the same surface of the plate-shaped body portion. In the first paragraph, The rear end of the above absorption chamber includes a waste collection and nucleic acid separation chamber in which the washing liquid is collected and the nucleic acid is discharged; A double-sided chip for nucleic acid extraction for PCR testing, characterized in that the nucleic acid separated through the waste collection and nucleic acid separation chamber is collected in the nucleic acid chamber. In the first paragraph, A washing liquid storage chamber is provided between the injection chamber and the absorption chamber to accommodate a washing liquid for washing the absorbent filled in the absorption chamber; The rear end of the above absorption chamber includes a waste collection and nucleic acid separation chamber in which the washing liquid is collected and the nucleic acid is discharged; A double-sided chip for nucleic acid extraction for PCR testing, characterized in that at least one of the washing liquid storage chamber, the absorption chamber, the waste collection and nucleic acid separation chamber is formed on at least one of one side of the body portion having a predetermined thickness and the other side thereof. In Article 12, The above washing liquid storage chamber and the above absorption chamber are formed on one side of the body portion, A double-sided chip for nucleic acid extraction for PCR testing, characterized in that the above-mentioned waste collection and nucleic acid separation chamber is formed on the other surface of the body part. In Article 12, A double-sided chip for nucleic acid extraction for PCR testing, characterized in that the body part further includes a tubular channel forming a channel through which a sample is transferred from the injection chamber to the nucleic acid chamber, and a valve for opening and closing the tubular channel is provided. In Article 14, The above tubular euro is, A first tube path provided between the injection chamber and the washing liquid storage chamber; A second tube path provided between the washing liquid storage chamber and the absorption chamber; and A third tube path provided between the above absorption chamber and the single waste waste collection and nucleic acid separation chamber; A double-sided chip for nucleic acid extraction for PCR testing, characterized in that the valve simultaneously opens or closes the first, second, and third tube channels.

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