PCR testing chip and PCR testing device including the same

Abstract

The present invention relates to a PCR testing chip and a PCR testing device including the same, and the PCR testing chip is characterized in that it includes: a body portion made of a light-transmitting material and having an inlet for introducing a sample; a reaction chamber having a first chamber portion recessed from the lower surface to the upper surface of the body portion and recessed from the lower surface to a first region, and a second chamber portion inside the first region, further recessed toward the upper surface, and containing a reaction reagent; and an inlet flow path connecting the inlet and the reaction chamber and guiding a sample to the reaction chamber. [Representative diagram] Figure 2

Description

[Technical field]

[0001] The present invention relates to a PCR (polymerase chain reaction) testing chip and a PCR testing device including the same, and to a PCR testing chip in which a reaction reagent that reacts with a sample is contained and fixed in a reaction chamber, thereby improving sensitivity when checking test results, and a PCR testing device including the same. [Background technology]

[0002] Polymerase chain reaction (PCR) is a method for amplifying specific target genetic material that one wishes to detect, and is a technique for amplifying large amounts of genetic material with identical base sequences from a small amount of genetic material. PCR is used to amplify human DNA to diagnose various types of genetic diseases, or to amplify the DNA of bacteria, viruses, and fungi to diagnose infectious diseases.

[0003] In general, PCR is carried out by repeating 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, and the primer is bound to the specific base sequence of the single-stranded DNA to form a partial DNA-primer complex; and (3) an extension step in which the sample solution is maintained at an active temperature of DNA polymerase, for example, 72°C, after the annealing step, and double-stranded DNA is formed based on the primer of the partial DNA-primer complex by DNA polymerase. By repeating the three steps several times, a target nucleic acid having a specific base sequence can be amplified exponentially.

[0004] To carry out such PCR, a PCR chip having a chamber capable of accommodating not only reagents and samples but also a sample solution, and a PCR device that heats and cools the PCR chip to induce an amplification reaction and measures the results are required. The PCR device is a specific means for realizing the amplification of a target nucleic acid, and various devices are currently being developed.

[0005] Conventional PCR only allows qualitative analysis of the amplified target nucleic acid using a separate electrophoresis after the reaction is completed, but recently, real-time PCR devices (i.e., rPCR devices) have been developed that use an optical detection system to detect the intensity of fluorescence proportional to the concentration of the amplified genetic material, allowing quantitative analysis of the target nucleic acid. In addition, a portable real-time PCR device has been developed that is a miniaturized version of the large PCR device used in laboratories and can perform PCR at the site where the sample is collected. The portable PCR device uses a PCR chip (or biochip, microchip) and is characterized by using a camera or digital camera instead of a large and complicated optical detection system.

[0006] The present invention relates to a PCR test chip that improves sensitivity when detecting the intensity of fluorescence emitted when a reagent reacts with a sample, and detects several target genes simultaneously. Summary of the Invention [Problem to be solved by the invention]

[0007] The present invention aims to provide a PCR testing chip and a PCR testing device including the same, in which a reaction reagent that reacts with a sample is contained and fixed in a reaction chamber, thereby improving sensitivity when checking test results. [Means for solving the problem]

[0008] According to one aspect of the present invention, there is provided a polymerase chain reaction (PCR) testing chip, comprising: a body portion made of a light-transmitting material and having an inlet for introducing a sample; a reaction chamber having a first chamber portion recessed from the lower surface to the upper surface of the body portion and recessed from the lower surface to a first region, and a second chamber portion inside the first region, recessed further toward the upper surface, and containing a reaction reagent; and an inlet flow channel connecting the inlet and the reaction chamber and guiding a sample to the reaction chamber.

[0009] It is also preferable that a plurality of reaction chambers are provided in the body portion.

[0010] It is also preferable that a plurality of reaction chambers are provided in the body portion, and that the reaction chambers are connected to each other by a connecting flow channel so that the sample flows in one direction.

[0011] Furthermore, it is desirable that the reaction chambers are arranged to form a first row portion in which the reaction chambers are arranged spaced apart from each other in a first direction, and a second row portion that is spaced apart from the first row portion, is parallel to the first direction, and is arranged between the chamber portions of the first row portion, and that the reaction chambers forming the first row portion and the reaction chambers forming the second row portion are arranged in a zigzag pattern with respect to each other.

[0012] Furthermore, when the reaction chamber is designated as the Nth chamber (N is a natural number) in order of proximity to the input port, it is desirable that the connecting flow path connecting the Nth chamber and the (N+1)th chamber includes a first flow path having one end connected to the Nth chamber, a second flow path having the other end connected to the (N+1)th chamber, and a direction change flow path connecting the first flow path and the second flow path and having a bent portion where the flow direction of the sample is changed.

[0013] It is also preferable that at least two bent portions are provided.

[0014] In addition, the reaction chambers are provided in a plurality of portions in the body portion and include a connecting channel connecting the reaction chambers to each other so that the sample flows in one direction. The lower surface of the body portion is provided with an inlet channel, a connecting channel, and a cover portion for closing the reaction chambers, the inlet channel and the connecting channel being groove-shaped and provided from the lower surface to the upper surface, and the cover portion preferably forms the bottom surface of the inlet channel, the connecting channel, and the reaction chamber.

[0015] Also, the cover is preferably disposed under the body and attached to the underside of the body in the form of a film so as to transmit heat from a heating device for carrying out PCR.

[0016] In addition, it is preferable that the depth of the connection channel recessed from the bottom surface of the body portion and the depth of the first chamber portion are the same.

[0017] In addition, the second chamber section preferably includes a first inclined flow path formed in a direction in which the sample flows in, and a second inclined flow path formed in a direction in which the sample flows out of the second chamber section. The first inclined flow path is formed to have a depth from the bottom surface of the body section that gradually increases as it approaches the inner surface of the second chamber section from a point spaced a predetermined distance from the second chamber section to the inner surface of the second chamber section, and the second inclined flow path is preferably formed opposite the first inclined flow path such that the depth from the bottom surface of the body section that gradually increases as it approaches the inner surface of the second chamber section from a point spaced a predetermined distance from the second chamber section to the inner surface of the second chamber section.

[0018] Also, it is preferable that the first and second inclined flow passages are curved and have a width that increases toward the inner circumferential surface of the second chamber portion.

[0019] Furthermore, it is preferable that, on the inlet side where the sample flows into the reaction chamber, the connecting flow path includes a first expansion section whose width gradually increases toward the reaction chamber, and, on the outlet side where the sample is discharged from the reaction chamber, the connecting flow path includes a second expansion section whose width gradually increases toward the reaction chamber.

[0020] In addition, it is desirable that at least two or more types of reaction reagents are provided so as to detect different target genes, and that different reaction reagents are contained in multiple reaction chambers, enabling multiple diagnosis of a single sample.

[0021] In addition, a PCR testing device according to another aspect of the present invention includes a main body: a body made of a light-transmitting material and having an inlet for introducing a sample; a plurality of reaction chambers having a first chamber section recessed from the bottom to the top of the body section, the first chamber section being recessed from the bottom to a first region, and a second chamber section inside the first region, recessed further toward the top, and containing a reaction reagent; an inflow flow path connecting the inlet and the reaction chambers and guiding a sample to the reaction chamber; and a connecting flow path connecting the plurality of reaction chambers to each other so that the sample flows in one direction; and a PCR testing chip attached to the main body: a heating device provided in the main body for heating the PCR testing chip; a light source provided in the main body for irradiating light to the PCR testing chip; a camera for capturing fluorescence emitted by the fluorescent dye when the reaction reagent containing a fluorescent dye reacts with the sample; and an analysis unit for analyzing the image captured by the camera.

[0022] Here, the reaction chambers are preferably arranged to form a first row portion in which the reaction chambers are spaced apart from each other in a first direction, and a second row portion that is spaced apart from the first row portion, parallel to the first direction, and arranged between the chamber portions of the first row portion, and the reaction chambers forming the first row portion and the reaction chambers forming the second row portion are preferably arranged in a zigzag pattern with respect to each other.

[0023] Here, when the reaction chamber is designated as the Nth chamber (N is a natural number) in the order adjacent to the input port, it is preferable that the connecting flow path connecting the Nth chamber and the (N+1)th chamber includes a first flow path having one end connected to the Nth chamber, a second flow path having the other end connected to the (N+1)th chamber, and a direction change flow path connecting the first flow path and the second flow path and having a bent portion where the flow direction of the sample is changed.

[0024] Here, a cover part for closing the inlet flow passage, the connecting flow passage, and the reaction chamber, which are provided in a groove shape from the bottom to the top, is attached to the bottom surface of the body part, and it is preferable that the cover part is provided in a film shape to transmit heat from a heating device and is attached to the bottom surface of the body part, and the cover part forms the bottom surface of the inlet flow passage, the connecting flow passage, and the reaction chamber.

[0025] Here, it is preferable that the second chamber section includes a first inclined flow path formed in a direction in which the sample flows in, and a second inclined flow path formed in a direction in which the sample flows out of the second chamber section, and that the first inclined flow path is formed to have a depth from the bottom surface of the body section that gradually increases as it approaches the inner surface of the second chamber section from a point spaced a predetermined distance from the second chamber section to the inner surface of the second chamber section, and that the second inclined flow path is formed opposite the first inclined flow path such that the depth from the bottom surface of the body section that gradually increases as it approaches the inner surface of the second chamber section from a point spaced a predetermined distance from the second chamber section to the inner surface of the second chamber section.

[0026] Here, it is preferable that, on the inlet side through which the sample flows into the reaction chamber, the connecting flow path includes a first expansion section whose width gradually increases toward the reaction chamber, and, on the outlet side through which the sample is discharged from the reaction chamber, the connecting flow path includes a second expansion section whose width gradually increases toward the reaction chamber. Effect of the Invention

[0027] The PCR testing chip and the PCR testing device including the same according to the embodiment of the present invention provide an effect of improving sensitivity when checking the test results by containing and fixing a reaction reagent that reacts with a sample in a reaction chamber.

[0028] Furthermore, according to the embodiment of the present invention, there is provided an advantage that a plurality of reaction chambers are provided, each of which is filled with various types of reaction reagents, and a plurality of target genes can be detected by a single sample injection.

[0029] In addition, by forming the reaction chamber in two stages and fixing the reaction reagent in the deeper chamber, the reaction reagent is captured and fixed. This provides the effect of concentrating the intensity of fluorescence during the reaction between the sample and the reaction reagent, improving visibility and improving the sensitivity and accuracy of detection.

[0030] In addition, the connecting flow path connecting the reaction chambers is provided with a bent portion, which effectively prevents the reaction reagent in one reaction chamber from migrating to an adjacent reaction chamber when the sample reacts with the reaction reagent, thereby improving the accuracy of the test results.

[0031] In addition, bubbles generated in the reaction chamber during sample injection can be pushed along the streamlined shape of the reaction chamber and removed.

[0032] In addition, the inlet side through which the sample flows into the reaction chamber and the outlet side through which the sample flows out of the reaction chamber are formed to be wider than the width of the connecting flow channel, which provides the effect that the sample can be easily distributed (distributed) to multiple reaction chambers. [Brief description of the drawings]

[0033] [Figure 1] FIG. 1 is a perspective view of a PCR testing chip according to an embodiment of the present invention. [Diagram 2] FIG. 2 is a bottom perspective view of FIG. 1. [Diagram 3] FIG. 2 is a bottom view of FIG. [Figure 4] FIG. 4 is an enlarged view of FIG. [Diagram 5] FIG. 13 is a diagram showing how the sample reacts with the reagent in the chamber, improving sensitivity. [Figure 6] FIG. 2 is a cross-sectional view of FIG. [Figure 7] FIG. 1 is a block diagram of a PCR device including a PCR testing chip. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0034] Hereinafter, various embodiments of the present invention will be described in conjunction with the accompanying drawings. Although various embodiments of the present invention can be modified in various ways and can have various embodiments, specific embodiments are illustrated in the drawings and related detailed descriptions are described. However, it should be understood that the various embodiments of the present invention are not limited to the specific embodiments, but include all modifications and / or equivalents or alternatives included in the spirit and technical scope of the various embodiments of the present invention. In conjunction with the description of the drawings, similar reference numerals are used for similar components.

[0035] The terms "comprise" or "may include" as used in the various embodiments of the present invention indicate the presence of the disclosed feature, operation, component, etc., but do not limit the inclusion of one or more additional features, operations, components, etc. Furthermore, in the various embodiments of the present invention, the terms "comprise" or "have" as specifying the presence of a feature, number, steps, operations, components, parts, or combinations thereof described in the specification, should be understood as not precluding the presence or addition of one or more other features, numbers, steps, operations, components, parts, or combinations thereof.

[0036] When a component is said to be "coupled" to another component, it should be understood that the component may be directly coupled to the other component, but there may be other components between the component and the other component. On the other hand, when a component is said to be "directly coupled" or "directly connected" to another component, it should be understood that there are no other components between the component and the other component.

[0037] The terms used in the various embodiments of the present invention are merely used to describe specific embodiments, and are not intended to limit the various embodiments of the present invention. A singular expression includes a plural expression unless otherwise clearly indicated in the context.

[0038] 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 the various embodiments of the present invention pertain.

[0039] Terms that are commonly used and defined in advance should be construed to have a meaning consistent with the meaning they have in the context of the relevant art, and should not be construed in an ideal or overly formal sense unless expressly defined in various embodiments of the present invention.

[0040] The present invention relates to a chip used in a polymerase chain reaction (PCR) device. The chip contains reaction reagents, and after a sample is injected into the chip, the chip is used to detect a target gene using a PCR device.

[0041] First, the term "PCR (polymerase chain reaction)" as used herein refers to a reaction that uses a thermostable DNA polymerase to amplify a specific target nucleic acid molecule. In addition to DNA polymerase, PCR may use a reaction mixture that contains primers (forward primer, reverse primer), which are oligonucleotides that can be hybridized specifically to the target nucleic acid, a deoxynucleotide triphosphate mixture (dNTP mixture), and divalent ions such as Mg2+.

[0042] "Primer" refers to an oligonucleotide or polynucleotide used to initiate a PCR reaction and hybridize to a template DNA in a complementary manner. The primers for the PCR reaction may be a pair of a forward primer (or a sense primer) selected from the same sense strand as the genetic coding direction of the nucleic acid molecule to be amplified, and a reverse primer (or an antisense primer) selected from the antisense strand complementary to the sense strand.

[0043] The term "sample" refers to the genetic material to be amplified, or a biological solution containing such genetic material. The term "reaction reagent" refers to a material for detecting the aforementioned target genetic material, and also includes primers and the like. The primers are a pair of primers having a length of 15 to 30 bp that can bind to both ends of a specific site of a target gene. The DNA polymerase used is an enzyme that does not lose its activity even at high temperatures of 90°C or higher.

[0044] DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Hereinafter, preferred embodiments of the present invention will be described in detail with reference to the accompanying drawings. A PCR testing chip 100 according to an embodiment of the present invention includes a body part 10, a reaction chamber 20 and an inlet channel 30. As shown in FIG.

[0045] The body part 10 is made of a light-transmitting material and has an inlet 11 for inserting a sample. The body part 10 is made of a light-transmitting material so that a fluorescent material can be detected by an optical detection device. According to this embodiment, the body part 10 is also made of a transparent polymeric material. The body part 10 is also rectangular, and protruding walls 12 are formed on opposing long sides of the rectangle so that the user can easily grasp it. Reinforcing protrusions 13 extending in the vertical direction are formed on the inside of the protruding walls 12. The reinforcing protrusions 13 prevent warping of the injection product during injection molding of the body part 10 and improve the rigidity of the body part 10. Meanwhile, a groove can be formed on the outer circumferential surface of the long side of the body part 10 instead of the protruding walls 12 so that the user can grasp the body part 10.

[0046] The reaction chamber 20 provides a space in which a sample and a reaction reagent react with each other, and includes a first chamber portion 21 and a second chamber portion 22 according to the present embodiment.

[0047] The first chamber 21 is recessed from the bottom surface of the body 10 toward the top surface. The first chamber 21 is recessed from the bottom surface of the body 10 to a depth of 0.3 mm to 0.5 mm. According to the present embodiment, the first chamber 21 is formed to a depth of 0.4 mm. If the depth of the first chamber 21 is less than 0.3 mm, the moving speed of the sample introduced therein is slow, and if it exceeds 0.5 mm, the efficiency of heat transfer to the reaction reagent is reduced. The first chamber 21 is recessed by an area corresponding to a predetermined first region. Overall, the first region of the first chamber 21 is formed in a circular shape, and an inlet through which the sample is introduced and an outlet through which the sample is discharged are formed.

[0048] The second chamber section 22 is formed inside the first region, with a step from the first region, and is further recessed toward the upper surface. That is, the second chamber section 22 is formed recessed with a step inside the first chamber section 21, so that the reaction chamber 20 is formed with a two-stage structure. The second chamber section 22 accommodates a reaction reagent. The second chamber section 22 is formed recessed from the lower surface of the body section 10 to a depth of 0.8 mm to 1.1 mm. It is preferable that the second chamber section 22 has a volume of 5 μl or more to effectively ensure the performance (luminescence) of the fluorescent material contained in the immobilized reagent. The depth of the second chamber section 22 is formed to 0.8 mm or more, making it easy to confirm the fluorescent material. If the depth of the second chamber section 22 is formed to a depth of 1.1 mm or more, the thickness of the body section 10 must be further reinforced, which has the disadvantage of reducing the heat transfer efficiency to the reaction reagent. According to this embodiment, the depth of the second chamber section 22 is formed to 1.0 mm.

[0049] According to this embodiment, the reaction reagent that reacts with the sample may include a primer, etc. The reaction reagent may also include a fluorescent dye. Here, it goes without saying that the fluorescent dye may be injected into the chip together with the sample in the PCR method. The fluorescent dye emits light in a specific wavelength band during the DNA extension (or amplification) step. At least two or more types of reaction reagents may be provided to detect different target genes, and different reaction reagents may be contained in multiple reaction chambers.

[0050] Specifically, according to this embodiment, a plurality of reaction chambers 20 are provided, and different reaction reagents are fixed in the reaction chambers 20, and different target genes are detected by one sample injection, making it possible to perform multiple diagnoses. For example, a reaction reagent for detecting a specific target gene can be accommodated and fixed in some of the plurality of second chambers 22. Then, reaction reagents for detecting other target genes can be accommodated and fixed in the other second chambers 22. At this time, each reaction reagent can be fixed in a state containing a fluorescent dye. Here, it goes without saying that the fluorescent dye can be injected into the chip together with the sample. The fluorescent dyes fixed together with the different reaction reagents or injected together with the sample can be the same dye, and when the fluorescent dye reacts with the target gene in each second chamber 22 and amplification is performed, the luminescence can be detected, and the reaction in the second chamber 22 can be identified. That is, a single wavelength is irradiated, making it possible to perform multiple diagnoses at one time.

[0051] The reaction reagent is fixed in the second chamber 22, and the chip with the reaction reagent fixed thereon can be transported to a place where a PCR test is required and used as needed. The reaction reagent can be fixed in the second chamber 22 by a method such as natural drying, vacuum drying, or freeze drying. The surface of the reaction reagent fixed in the second chamber 22 can be coated with a coating agent having waterproof and adhesive properties. The coating agent can be fixed to the reagent (primer, probe, premix) using a biopolymer material that does not affect the reagent properties or PCR results. The coating agent can have a property of maintaining a solid state at room temperature and melting at a certain temperature or higher. For example, the coating agent can melt at 45° C. or higher. When the chip with the sample injected therein is heated during the PCR test, the coating agent can melt and react with the sample and the reaction reagent. The coating agent can be any one of wax, agarose, paraffin, collagen, chitosan, and gelatin, or a mixture thereof.

[0052] Here, it goes without saying that the coating agent may be mixed with the reaction reagent and then accommodated and fixed in the second chamber 22. Even in such a case, the reaction reagent accommodated in the second chamber 22 maintains a solid state at room temperature, and when the chip into which the sample is injected is heated during the PCR test, the coating agent may melt and react with the sample.

[0053] The second chamber section 22 is provided with a first inclined flow channel 221 formed in a direction in which the sample is introduced, and a second inclined flow channel 222 formed in a direction in which the sample is discharged from the second chamber section 22. The first inclined flow channel 221 is a flow channel that is connected from a point separated by a predetermined distance from the second chamber section 22 to the inner peripheral surface of the second chamber section 22, and is formed so that the depth from the lower surface of the body section 10 becomes gradually deeper as it approaches the inner peripheral surface of the second chamber section 22. The second inclined flow channel 222 is formed in the same shape as the first inclined flow channel 221. That is, the second inclined flow channel 222 is a flow channel that is connected from a point separated by a predetermined distance from the second chamber section 22 to the inner peripheral surface of the second chamber section 22, and is formed at a position symmetrical to the first inclined flow channel 221, and is formed so that the depth from the lower surface of the body section 10 becomes gradually deeper as it approaches the inner peripheral surface of the second chamber section 22.

[0054] 4, the first inclined flow channel 221 is formed so that its width increases toward the inner circumferential surface of the second chamber section 22 and is curved. That is, the left and right side surfaces of the first inclined flow channel 221 are formed so that their width increases along the sample inflow direction and are curved near the portion connected to the second chamber section 22. Similarly to the first inclined flow channel 221, the second inclined flow channel 222 is formed so that its width increases toward the inner circumferential surface of the second chamber section 22 and is curved. The second inclined flow channel 222 is disposed opposite the first inclined flow channel 221 and is formed so that the widths of the left and right side surfaces increase toward the inner circumferential surface of the second chamber section 22 and are curved.

[0055] The inlet flow path 30 is provided to connect the inlet 11 and the reaction chamber 20 and to guide the sample to the reaction chamber 20. According to the present embodiment, the inlet 11 is provided to protrude upward from the upper surface of the body part 10. Referring to FIG. 1, two protrusions protruding upward are formed on the upper surface of the body part 10, and the protrusions are provided to penetrate the body part 10 in the vertical direction. One of the two protrusions is also used as the inlet 11 for introducing the sample. The remaining protrusion communicates the flow path with the outside air when the sample is injected through the inlet 11, so that the sample flows along the flow path formed in the body part 10.

[0056] According to this embodiment, a plurality of reaction chambers 20 are provided in the body portion 10, and the reaction chambers 20 are connected to each other by a connecting flow channel 40 so that the sample flows in one direction.

[0057] According to this embodiment, as shown in FIG. 2 and FIG. 3, the reaction chambers 20 include a first row and a second row. In the first row, the reaction chambers 20 are arranged spaced apart from each other in a first direction. The reaction chambers 20 arranged in the first row are arranged on an imaginary straight line extending in one direction. The second row is spaced apart from the first row, parallel to the first direction, and arranged between the reaction chambers 20 in the first row. The reaction chambers 20 arranged in the second row are arranged on an imaginary straight line extending in one direction, similar to the first row, and the imaginary straight lines on which the first row and the second row are arranged are parallel to each other. With such a structure, the reaction chambers 20 forming the first row and the reaction chambers 20 forming the second row are arranged in a zigzag pattern. According to this embodiment, the first row includes seven reaction chambers 20, and the second row includes six reaction chambers 20, forming a total of 13 reaction chambers 20. As described above, the same or different reaction reagents can be loaded into each reaction chamber 20. According to this embodiment, 13 or less reaction reagents can be immobilized in the reaction chambers 20, and multiple diagnoses can be performed by distinguishing target genes according to the number of immobilized reaction reagents. However, the number of reaction chambers 20 is not limited to the above example, and various numbers and intervals can be formed.

[0058] The connecting channel 40 is provided to connect the multiple reaction chambers 20 to each other.

[0059] If the reaction chambers 20 are designated as N-th chambers (N is a natural number) in order of their proximity to the inlet 11, the connecting channel 40 connects the N-th chamber to the (N+1)-th chamber. Here, "in order of their proximity to the inlet 11" refers to the order in which the sample passes when it passes through the reaction chambers 20 in sequence via the connecting channel 40, starting from the reaction chamber 20 to which it first reaches via the inflow channel 30 after being injected through the inlet 11. According to this embodiment, the connecting channel 40 includes a first channel 41, a second channel 42, and a direction change channel 43.

[0060] The first flow path 41 has one end connected to the Nth chamber, and the other end extending toward the (N+1)th chamber. The second flow path 42 has the other end connected to the (N+1)th chamber, and one end extending toward the Nth chamber. The direction change flow path 43 connects the first flow path and the second flow path. One end of the direction change flow path 43 is connected to the other end of the first flow path 41, and the other end is connected to one end of the second flow path 42. According to this embodiment, the direction change flow path 43 includes a bent portion 431 that changes the flow direction of the sample. According to this embodiment, at least two bent portions 431 are provided.

[0061] As shown in FIG. 4, the bending portion 431 according to the present embodiment includes a first bending portion 4311 and a second bending portion 4312. The first bending portion 4311 changes the flow direction of the sample by about 90 degrees, and the second bending portion 4312 changes the flow direction of the sample by 180 degrees. According to the present embodiment, the sample flowing out from the Nth chamber changes its direction by about 90 degrees through the first bending portion 4311, then passes through the second bending portion 4312 three times in succession, and then changes its direction by about 90 degrees through the first bending portion 4311 again to be introduced into the (N+1)th chamber. According to the present embodiment, the bending portion 431 is formed with two first bending portions 4311 and three second bending portions 4312, but the number of the first bending portions 4311 and the second bending portions 4312 is not limited thereto and may be formed in various ways.

[0062] The bend portion 431 prevents the sample from moving between adjacent reaction chambers 20 when the sample is injected into any of the reaction chambers 20 provided in the body portion 10 and reacts with the reagent, thereby ensuring reliability by preventing the reaction results of each reaction chamber 20 from being mixed with the reaction results of other reaction chambers 20.

[0063] According to this embodiment, the connecting flow path 40 includes a first expansion portion 23 and a second expansion portion 24 .

[0064] The first expansion section 23 is a section whose width gradually increases from an inlet side through which a sample is introduced into the reaction chamber 20 toward the reaction chamber 20. The first expansion section 23 is formed in a streamlined shape, and when the sample flows into the reaction chamber 20, it slows down along the section where the width increases and fills the internal space of the reaction chamber 20.

[0065] The second expansion section 24 is a section whose width gradually increases from the outlet side, through which the sample is discharged from the reaction chamber 20, toward the reaction chamber 20. The second expansion section 24 is formed symmetrically opposite the first expansion section 23, and when the sample flows out of the reaction chamber 20, the width of the second expansion section 24 decreases as it moves away from the reaction chamber 20, and the flow speed of the sample increases, allowing the sample to easily move to another adjacent reaction chamber 20.

[0066] According to the present embodiment, the depth of the connecting passage 40 recessed from the bottom surface of the body portion 10 is formed to be equal to the depth of the first chamber portion 21. As shown in Fig. 4, the second chamber portion 22 provided inside the first chamber portion 21 is formed to be deeper than the connecting passage 40, and the step portion between the first chamber portion 21 and the second chamber portion 22 is formed to be the same depth as the connecting passage 40.

[0067] According to this embodiment, the cover portion 50 is coupled to the lower surface of the body portion 10 .

[0068] The cover part 50 is provided to close the inflow channel 30, the connecting channel 40, and the reaction chamber 20, which are formed in a groove shape from the lower surface to the upper surface of the body part 10. The cover part 50 is attached to the bottom surface of the body part 10 in a state in which a sample is accommodated and fixed in the reaction chamber 20. The cover part 50 forms the bottom surfaces of the inflow channel 30, the connecting channel 40, and the reaction chamber 20. That is, the sample injected through the inlet 11 is transferred to the reaction chamber 20 through the inflow channel 30 and the connecting channel 40, and at this time, the bottom surfaces of the inflow channel 30, the connecting channel 40, and the reaction chamber 20 are formed by the cover part 50.

[0069] According to this embodiment, the cover unit 50 is disposed under the body unit 10, and is provided in the form of a film so that heat can be transferred from the heating device 120 for PCR, and is attached to the underside of the body unit 10. The cover unit 50 is made of a thin film, and can easily transfer heat from the heating device 120, and can be made of a metal having excellent heat transfer efficiency or a polymeric material containing nanocarbon. The sample moves automatically through the inlet channel 30, the connecting channel 40, and the reaction chamber 20 without using any power.

[0070] The action and effect of the PCR testing chip 100 according to the configuration will be described in detail below.

[0071] As shown in FIG. 2, the inlet channel 30, the reaction channel, and the reaction chamber 20 are formed in a groove shape on the lower surface of the body part 10 according to the embodiment of the present invention. First, a reaction reagent is fixed in the second chamber part 22 of the reaction chamber 20. The reaction reagent may be fixed in the reaction chamber 20, where the reagents for detecting different target genes are different. The lower surface of the body part 10 is sealed by the cover part 50 in a film form. The cover part 50 closes the inlet channel 30, the connection channel 40, and the reaction chamber 20. According to the present embodiment, an adhesive region is provided on the edge of the inlet channel 30, the connection channel 40, and the reaction chamber 20 so that the cover part 50 in a film form can be attached. The adhesive region is formed wider than the width of the inlet channel 30 and the connection channel 40, so that sample leakage can be prevented when the sample is injected. The adhesive region may be formed along the edge of the inlet channel 30, the connection channel 40, and the reaction chamber 20 with a width of 0.8 mm to 1.0 mm. If the width of the adhesive region is less than 0.8 mm, the adhesive strength of the cover part 50 will be poor, and if it exceeds 1.0 mm, the spatial arrangement of the reaction chamber 20 will be restricted.

[0072] The reaction reagent may be fixed in the reaction chamber 20 together with a coating agent. The chip on which the reaction reagent is fixed is stored separately, and the chip according to the embodiment of the present invention is transported to a site where a PCR test is to be performed, where a sample is injected through the inlet 11 and the test is performed through a PCR device. Thus, the PCR test chip 100 according to the present invention allows the reaction reagent to be fixed in advance, quickly performing a PCR test, and allows multiple reaction reagents to be fixed in advance, enabling multiple tests.

[0073] A sample is injected through the inlet 11. Two protrusions are formed on the top surface of the body part 10, and either one of them can be used as the inlet 11. When a sample is injected into the selected inlet 11, the sample passes through the inlet channel 30 and the connecting channel 40 and fills the reaction chamber 20 provided in the body part 10. The remaining inlet 11 communicates with the outside air, and moves the sample itself along a narrow channel.

[0074] The sample is introduced into the reaction chamber 20 along the inlet flow channel 30, and then into the reaction chambers 20 arranged in sequence. The sample flows out from the Nth chamber along the first flow channel 41. When the sample leaves the Nth chamber, the moving speed of the sample increases as it passes through the second expansion section 24, and the sample easily moves to the (N+1)th chamber. The sample passes through the bent section 431 of the direction change flow channel 43, and the flow channel is changed as the sample advances. The bent section 431 is provided between the Nth chamber and the (N+1)th chamber, and forms a pressure boundary so that the sample cannot move to the adjacent chambers if it reacts with a reaction reagent. The sample that has passed through the direction change flow channel 43 is introduced into the (N+1)th chamber via the second flow channel 42. According to this embodiment, the sample flows from the first chamber section 21 to the thirteenth chamber along the above-mentioned route.

[0075] Furthermore, when a sample is introduced into the Nth chamber, the sample also flows along the first inclined flow channel 221 while entering the inlet. The first inclined flow channel 221 is formed so that its depth increases toward the second chamber portion 22, the width between the side portions increases, and the channel is curved, so that the sample effectively comes into contact with the reaction reagent fixed in the second chamber portion 22.

[0076] In addition, the reaction chamber 20 according to the present embodiment is formed in two stages by a first chamber part and a second chamber part, and a reaction reagent is fixed in the second chamber part 22, so that bubbles are generated in the process of the reaction reagent expanding upon contact with the sample, and are collected in the second chamber part 22. The sample entering the second chamber part 22 along the first inclined channel 221 can push the bubbles toward the second inclined channel 222 and remove them. Since the inner periphery of the second chamber part 22 is circular, the sample moves along the streamlined inner periphery and the bubbles are effectively removed. By removing the bubbles, the reaction result between the sample and the reaction reagent can be obtained accurately. In addition, the first inclined channel 221 and the second inclined channel 222 are provided to remove PCR inhibitors such as bubbles generated inside the reaction chamber 20 when the reaction reagent is dispensed.

[0077] 6, according to this embodiment, the reaction reagent is fixed in the second chamber 22, so that the fluorescent material is concentrated in a limited space, and the sensitivity to the reaction result during image acquisition and analysis through a PCR device can be improved, thereby improving the accuracy of diagnosis. In addition, since the reaction reagent reacts with the sample while being fixed in the well in the second chamber 22, the movement of the reaction sample between the reaction chambers 20 is suppressed, and the reaction samples reacted with the sample do not mix with each other, so that the reliability of the reaction result can be further improved.

[0078] According to another aspect of the present invention, a PCR device including the PCR testing chip 100 is provided.

[0079] The PCR device according to this embodiment is equipment for performing PCR by mounting the above-mentioned PCR testing chip 100 thereon, and as shown in FIG. 7, includes a main body 110 on which the PCR testing chip 100 is mounted, a heating means provided inside the main body 110 and providing heat to the chip 100, a light source 130 for irradiating light to the chip 100, a camera 140 for capturing fluorescence generated when the reaction reagent and the sample react with each other, and an analysis unit 150 for analyzing the image captured by the camera 140.

[0080] The main body 110 includes a mounting part on which the chip 100 is mounted. The main body 110 is provided with an operation panel for performing PCR, and the operation panel is provided with a number of input buttons to perform functions such as starting and ending a PCR test and operating the heating device 120. The mounting part protrudes from the main body 110 and moves to a position where the chip 100 can be inserted, and after the chip 100 is placed on the mounting part, it can be further moved inside the main body 110. It goes without saying that the shape of the mounting part is not limited to that described above.

[0081] The heating means is provided to heat the chip 100. The heating means is provided under the chip 100 and heats the reaction reagent and the sample to a temperature required for carrying out a PCR test. At this time, the heat of the heating means is transferred to the reaction reagent and the sample via the film-shaped cover part 50, enabling rapid heating.

[0082] The light source 130 is provided to irradiate light to the chip 100. The light source 130 may be provided by an LED provided in the body 110. A plurality of LEDs may be provided, and the light provided by the LEDs may be a single wavelength having the same wavelength or light having different wavelengths.

[0083] The camera 140 is provided to capture the fluorescence emitted by the fluorescent dye in response to the light irradiated from the light source 130. The camera 140 is disposed above the chip 100 and captures images of multiple reaction chambers 20 at one time. The camera 140 can capture fluorescent images at regular intervals above the chip 100.

[0084] The analysis unit 150 analyzes the image captured by the camera 140. The analysis unit 150 analyzes the hue of the image by comparing it with predetermined reference data. The predetermined reference data is data in which various target genes or diseases are matched in advance according to the type of color (RGB ratio) and intensity of emitted fluorescent light, and the analysis unit 150 can calculate the RGB ratio of the image captured by the camera 140 using an RGB analysis program and compare it with the predetermined data. The result value by the analysis unit 150 can be displayed on the display unit 160 and provided to the user.

[0085] Through such a process, the present invention enables multiplex testing by detecting various kinds of target genes with one sample injection. The PCR device including the PCR testing chip according to the present invention directly employs the PCR testing chip 100 described above, and therefore can provide the same functions and effects as the PCR testing chip 100.

[0086] The present invention has been described in detail above with reference to preferred embodiments, but the present invention is not limited to the above embodiments, and various modifications may be made without departing from the scope of the present invention.

Claims

1. a body portion made of a light-transmitting material and having an inlet through which a sample is inserted; a reaction chamber including a first chamber portion recessed from the lower surface of the body portion to an upper surface of the body portion in a first region of the lower surface of the body portion, and a second chamber portion recessed further from the first chamber portion to the upper surface of the body portion within the first region, in which a reaction reagent is contained; an inlet channel connecting the inlet and the reaction chamber and guiding the sample to the reaction chamber,

2. 2. The PCR testing chip according to claim 1, wherein a plurality of said reaction chambers are provided in said body portion.

3. The reaction chamber is provided in a plurality of parts in the body portion, 2. The PCR testing chip according to claim 1, wherein the reaction chambers are connected to each other by a connecting channel so that the sample flows in one direction.

4. The reaction chamber comprises: The reaction chambers are arranged to form a first row in which the reaction chambers are spaced apart from each other in a first direction, and a second row that is spaced apart from the first row, parallel to the first direction, and disposed between the chambers in the first row; 2. The PCR testing chip according to claim 1, wherein the reaction chambers forming the first row and the reaction chambers forming the second row are arranged in a zigzag pattern.

5. When the reaction chamber is an N-th chamber (N is a natural number) in the order adjacent to the inlet, The connecting flow path connecting the Nth chamber and the (N+1)th chamber is A first flow path having one end connected to the Nth chamber; a second flow path having the other end connected to the (N+1)th chamber; 4. The PCR testing chip according to claim 3, further comprising: a direction-changing flow channel that connects the first flow channel and the second flow channel and has a bent portion that changes a flow direction of the sample.

6. 6. The PCR testing chip according to claim 5, wherein at least two of the bent portions are provided.

7. The reaction chamber is provided in a plurality of parts in the body portion, The PCR testing chip is a connecting channel connecting the reaction chambers to each other so that the sample flows in one direction; a cover part for closing the inlet channel, the connection channel, and the reaction chamber, the inlet channel being formed in a groove shape from the lower surface of the body part to the upper surface of the body part, 2. The PCR testing chip according to claim 1, wherein the cover part forms the bottom surface of the inlet channel, the connecting channel, and the reaction chamber.

8. The PCR testing chip according to claim 7, characterized in that the cover part is disposed under the body part, formed in a film shape to transmit heat from a heating device for performing a PCR test, and attached to the underside of the body part.

9. 8. The PCR testing chip according to claim 7, wherein the depth of the connection channel recessed from the lower surface of the body portion is equal to the depth of the first chamber portion.

10. the second chamber includes a first inclined flow path formed in a direction in which the sample is introduced, and a second inclined flow path formed in a direction in which the sample is discharged from the second chamber, the first inclined flow passage is formed such that a depth from the bottom surface of the body portion gradually increases from a point spaced a predetermined distance from the second chamber portion to an inner peripheral surface of the second chamber portion as the first inclined flow passage approaches the inner peripheral surface, 2. The PCR testing chip according to claim 1, wherein the second inclined flow path is formed opposite to the first inclined flow path, and is formed such that a depth of the second inclined flow path from the bottom surface of the body portion gradually increases as the second inclined flow path approaches the inner circumferential surface of the second chamber portion from a point spaced a predetermined distance from the second chamber portion to the inner circumferential surface of the second chamber portion.

11. 11. The PCR testing chip according to claim 10, wherein the first inclined flow channel and the second inclined flow channel are formed so that their widths gradually increase toward the inner circumferential surface of the second chamber portion and are curved.

12. At an inlet side through which the sample is introduced into the reaction chamber, the connecting channel has a first expansion part whose width gradually increases toward the reaction chamber, 4. The PCR testing chip according to claim 3, wherein the connecting channel has a second expansion portion, the width of which gradually increases as the connecting channel approaches the reaction chamber, at a side of an outlet through which the sample is discharged from the reaction chamber.

13. The reaction reagent includes at least two or more types of reaction reagents so as to detect different target genes from each other, 3. The PCR testing chip according to claim 2, wherein different kinds of reaction reagents are accommodated in the reaction chambers to enable multiple diagnoses for one sample.

14. A main body portion, a chip for PCR testing comprising: a body portion made of a light-transmitting material and having an inlet through which a sample is introduced; a first chamber portion formed in a first region of a lower surface of the body portion so as to be recessed from the lower surface to an upper surface of the body portion; and a plurality of reaction chambers having a second chamber portion formed in the first region so as to be further recessed from the first chamber portion to the upper surface of the body portion and for containing a reaction reagent; an inflow channel connecting the inlet and the reaction chambers and guiding the sample to the reaction chambers; and a connection channel connecting the plurality of reaction chambers to each other so that the sample flows in one direction, the chip being attached to the main body portion; a heating device provided in the main body for heating the PCR testing chip; A light source provided in the main body for irradiating light onto the PCR testing chip; a camera that captures the fluorescence emitted by the fluorescent dye when the reaction reagent containing the fluorescent dye reacts with the sample; An analysis unit that analyzes the image captured by the camera.

15. The reaction chamber comprises: The reaction chambers are arranged to form a first row in which the reaction chambers are spaced apart from each other in a first direction, and a second row that is spaced apart from the first row, parallel to the first direction, and disposed between the chambers in the first row; 15. The PCR testing device according to claim 14, wherein the reaction chambers forming the first row and the reaction chambers forming the second row are arranged in a zigzag pattern relative to each other.

16. When the reaction chamber is an N-th chamber (N is a natural number) in the order adjacent to the inlet, The connecting flow path connecting the Nth chamber and the (N+1)th chamber is A first flow path having one end connected to the Nth chamber; a second flow path having the other end connected to the (N+1)th chamber; 15. The PCR testing device according to claim 14, further comprising: a direction-changing flow channel that connects the first flow channel and the second flow channel and has a bent portion that changes a flow direction of the sample.

17. a cover part attached to the lower surface of the body part and formed in a groove shape from the lower surface of the body part to the upper surface of the body part to close the inlet channel, the connection channel, and the reaction chamber; the cover portion is formed in a film shape so as to transmit heat from the heating device, and is attached to the lower surface of the body portion; The PCR testing device according to claim 14, wherein the cover part forms the bottom surface of the inlet channel, the connecting channel and the reaction chamber.

18. the second chamber includes a first inclined flow path formed in a direction in which the sample is introduced, and a second inclined flow path formed in a direction in which the sample is discharged from the second chamber, the first inclined flow passage is formed such that a depth from the bottom surface of the body portion gradually increases from a point spaced a predetermined distance from the second chamber portion to an inner peripheral surface of the second chamber portion as the first inclined flow passage approaches the inner peripheral surface, The PCR testing device according to claim 14, characterized in that the second inclined flow path is formed opposite to the first inclined flow path, and is formed such that its depth from the bottom surface of the body portion gradually increases as it approaches the inner surface from a point a predetermined distance away from the second chamber portion to the inner surface of the second chamber portion.

19. At an inlet side through which the sample is introduced into the reaction chamber, the connecting channel has a first expansion part whose width gradually increases toward the reaction chamber, 15. The PCR testing device of claim 14, wherein the connecting channel has a second expansion portion, the width of which gradually increases as the connecting channel approaches the reaction chamber, at a side of an outlet through which the sample is discharged from the reaction chamber.

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