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 relates 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 equipped with the same. [Background technology]

[0002] Polymerase chain reaction (PCR) is a method for amplifying specific target genetic material, enabling the amplification of large amounts of genetic material with identical base sequences from small amounts of genetic material. PCR is used to amplify human DNA to diagnose various genetic disorders, and is also used to diagnose infectious diseases by amplifying the DNA of bacteria, viruses, and fungi.

[0003] Typically, PCR involves the repeated execution of three steps: (1) a denaturing step in which a sample solution containing double-stranded DNA is heated to a specific temperature, e.g., about 95°C, to separate the double-stranded DNA into single-stranded DNA; (2) an annealing step in which, after the denaturing step, oligonucleotide primers having a sequence complementary to the specific base sequence to be amplified are added to the sample solution and cooled together with the separated single-stranded DNA to a specific temperature, e.g., 55°C, to allow the primers to bind to the specific base sequence of the single-stranded DNA, forming a partial DNA-primer complex; and (3) an extension (or amplification) step in which, after the annealing step, the sample solution is maintained at a temperature at which DNA polymerase is active, e.g., 72°C, to allow the DNA polymerase to form double-stranded DNA based on the primers in the partial DNA-primer complex. Repeating these three steps several times allows for exponential amplification of a target nucleic acid having a specific base sequence.

[0004] To perform such PCR, a PCR chip equipped with 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 target nucleic acids, and various devices are currently being developed.

[0005] Conventional PCR only allows for qualitative analysis of the amplified target nucleic acid after the reaction is completed using separate electrophoresis. However, real-time PCR devices (i.e., rPCR devices) have recently been developed that use an optical detection system to detect fluorescence intensity, which is proportional to the concentration of amplified genetic material, allowing for quantitative analysis of the target nucleic acid. Additionally, portable real-time PCR devices have been developed that are a miniaturization of the large PCR devices used in laboratories, allowing PCR to be performed at the site where the sample is collected. These portable PCR devices use a PCR chip (or biochip or microchip) and a camera or digital camera instead of a large and complex optical detection system.

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

[0007] The present invention aims to provide 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. [Means for solving the problem]

[0008] A polymerase chain reaction (PCR) testing chip according to one aspect of the present invention 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 bottom to the top of the body portion and recessed from the bottom to a first region; and a second chamber portion inside the first region, recessed further toward the top, with a step from the first region, for accommodating a reaction reagent; and an inlet channel connecting the inlet and the reaction chamber and guiding the sample to the reaction chamber.

[0009] It is also desirable that a plurality of reaction chambers be 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 one another by connecting channels so that the sample flows in one direction.

[0011] Furthermore, the reaction chambers are preferably arranged to form a first row in which the reaction chambers are spaced apart from one another in a first direction, and a second row that is spaced apart from the first row, parallel to the first direction, and arranged between the chambers of the first row, and the reaction chambers that make up the first row and the reaction chambers that make up the second row are preferably arranged in a zigzag pattern.

[0012] Furthermore, when the reaction chamber is designated as the Nth chamber (N is a natural number) in order of proximity to the inlet, the connecting flow path connecting the Nth chamber and the (N+1)th chamber preferably 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-changing 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 desirable that at least two bent portions are provided.

[0014] In addition, the reaction chambers are provided in a plurality in the body portion and include a connecting channel that connects 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 that closes the reaction chambers, which are groove-shaped and are provided from the lower surface to the upper surface, and the cover preferably forms the inlet channel, the connecting channel, and the bottom surface of the reaction chambers.

[0015] The cover is preferably disposed under the body and attached to the underside of the body in the form of a film to transmit heat from a heating device for PCR.

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

[0017] 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 so that its depth from the bottom surface of the body section 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. The second inclined flow path is preferably formed opposite the first inclined flow path so that its depth from the bottom surface of the body section 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] Preferably, the first and second inclined flow paths are curved and have a width that increases as they approach the inner circumferential surface of the second chamber portion.

[0019] Furthermore, it is desirable that the connecting flow path includes a first expansion section at the inlet side where the sample flows into the reaction chamber, the width of which gradually increases as the connecting flow path approaches the reaction chamber, and a second expansion section at the outlet side where the sample is discharged from the reaction chamber, the width of which gradually increases as the connecting flow path approaches the reaction chamber.

[0020] Furthermore, 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 diagnoses to be performed on 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 each having a first chamber portion recessed from the bottom to the top of the body portion and recessed from the bottom to a first region, and a second chamber portion inside the first region, recessed further toward the top, and accommodating a reaction reagent; an inlet flow path connecting the inlet and the reaction chambers and guiding the 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; a PCR testing chip attached to the main body: a heating device attached to the main body and heating the PCR testing chip; a light source attached to the main body and irradiating light onto the PCR testing chip; a camera for capturing fluorescence emitted by the fluorescent dye when the reaction reagent containing the 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 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 arranged between the chambers in the first row, and the reaction chambers in the first row and the reaction chambers in the second row are preferably arranged in a zigzag pattern.

[0023] Here, when the reaction chamber is designated as the Nth chamber (N is a natural number) in the order of its proximity to the inlet, the connecting flow path connecting the Nth chamber and the (N+1)th chamber preferably 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-changing 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 channel, the connecting channel, and the reaction chamber, which are groove-shaped from the bottom to the top, is attached to the bottom surface of the body part, and the cover part is preferably provided in the form of a film to transfer heat from a heating device and attached to the bottom surface of the body part, and the cover part preferably forms the bottom surface of the inlet channel, the connecting channel, and the reaction chamber.

[0025] Here, the second chamber section preferably includes a first inclined flow channel formed in a direction in which the sample flows in and a second inclined flow channel formed in a direction in which the sample flows out of the second chamber section. The first inclined flow channel is preferably formed so that its depth from the bottom surface of the body section 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 channel is preferably formed opposite the first inclined flow channel so that its depth from the bottom surface of the body section 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 the connecting flow path includes a first expansion section at the inlet side through which the sample flows into the reaction chamber, the width of which gradually increases as the connecting flow path approaches the reaction chamber, and a second expansion section at the outlet side through which the sample is discharged from the reaction chamber, the width of which gradually increases as the connecting flow path approaches the reaction chamber. [Effects 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 the effect of improving sensitivity when checking 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, which can be filled with various types of reaction reagents, and multiple target genes can be detected by a single sample injection.

[0029] In addition, by forming the reaction chamber in two stages and immobilizing the reaction reagent in the deeper chamber, the reaction reagent is captured and immobilized. This allows the fluorescent intensity to be concentrated 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 section, which effectively prevents the reaction reagent in one reaction chamber from moving to an adjacent reaction chamber when the sample and reaction reagent react, 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 wider than the width of the connecting channel, which provides the effect of easily distributing (diffusing) the sample to multiple reaction chambers. [Brief explanation of the drawings]

[0033] [Figure 1] 1 is a perspective view of a PCR testing chip according to an embodiment of the present invention. [Figure 2] FIG. 2 is a bottom perspective view of FIG. 1. [Figure 3] FIG. 2 is a bottom view of FIG. 1. [Figure 4] FIG. 4 is an enlarged view of FIG. [Figure 5] FIG. 10 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. 1. [Figure 7] FIG. 1 is a block diagram of a PCR device including a PCR testing chip. DETAILED DESCRIPTION OF THE INVENTION

[0034] Various embodiments of the present invention will be described below with reference to the accompanying drawings. Although various modifications and variations of the present invention are possible, specific embodiments are illustrated in the drawings and described in the related detailed description. However, this does not limit the various embodiments of the present invention to specific embodiments, and it should be understood that the present invention includes all modifications, equivalents, or alternatives that fall within the spirit and technical scope of the various embodiments of the present invention. Similar reference numerals are used for similar components in connection with the description of the drawings.

[0035] The terms "comprise" or "may comprise" as used in various embodiments of the present invention indicate the presence of the disclosed feature, operation, or component, but do not limit the presence of one or more additional features, operations, or components. Furthermore, in various embodiments of the present invention, the terms "comprise" or "have" as specifying the presence of a specified feature, number, step, operation, component, part, or combination thereof, and should not be understood to preclude the presence or addition of one or more other features, numbers, steps, operations, components, parts, or combinations thereof.

[0036] When a component is referred to as being "coupled" to another component, it should be understood that the component may be directly coupled to the other component, but that there may be other components between the component and the other component. On the other hand, when a component is referred to as being "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 particular embodiments and are not intended to limit the various embodiments of the present invention. The singular expressions include the plural expressions unless otherwise clearly indicated in the context.

[0038] Unless otherwise defined, all terms used herein, including technical and 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 commonly used and predefined 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 idealized 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, as used herein, the term "polymerase chain reaction (PCR)" refers to a reaction that uses a thermostable DNA polymerase to amplify a specific target nucleic acid molecule. PCR may use a reaction mixture that contains, in addition to the DNA polymerase, primers (forward primer, reverse primer) that are oligonucleotides that can hybridize 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. Primers for PCR reactions may be a pair of a forward primer (or sense primer) selected from the sense strand that is identical to the genetic coding direction of the nucleic acid molecule to be amplified, and a reverse primer (or antisense primer) selected from the antisense strand that is 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 reagent for detecting the target genetic material and includes primers. The primers may consist of a 15-30 bp primer pair capable of binding to both ends of a specific site in the target gene. The DNA polymerase used is an enzyme that does not lose its activity even at temperatures above 90°C.

[0044] DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Preferred embodiments of the present invention will now 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 10, a reaction chamber 20, and an inlet channel 30. As shown in FIG.

[0045] The body 10 is made of a light-transmitting material and has an inlet 11 for introducing a sample. The body 10 is made of a light-transmitting material so that the fluorescent material can be detected by an optical detection device. According to this embodiment, the body 10 is also made of a transparent polymeric material. The body 10 may be rectangular, and protruding walls 12 are formed on opposing long sides of the rectangle to make it easier for the user to grip. 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 extruded material during injection molding of the body 10 and improve the rigidity of the body 10. Alternatively, grooves may be formed on the outer circumferential surfaces of the long sides of the body 10 in place of the protruding walls 12 to allow the user to grip the body 10.

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

[0047] The first chamber 21 is recessed from the bottom to the top of the body 10. The first chamber 21 is recessed from the bottom 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 movement speed of the introduced sample becomes slow, while if it exceeds 0.5 mm, the efficiency of heat transfer to the reaction reagent decreases. 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 has an inlet through which the sample is introduced and an outlet through which the sample exits.

[0048] The second chamber portion 22 is recessed from the first region toward the upper surface, forming a step with the first region. That is, the second chamber portion 22 is recessed from the first chamber portion 21, forming a two-tiered reaction chamber 20. The second chamber portion 22 accommodates a reaction reagent. The second chamber portion 22 is recessed from the bottom surface of the body portion 10 to a depth of 0.8 mm to 1.1 mm. The second chamber portion 22 preferably has a volume of 5 μl or more to effectively ensure the performance (luminescence) of the fluorescent substance contained in the immobilized reagent. The depth of the second chamber portion 22 is 0.8 mm or more, making it easy to identify the fluorescent substance. If the depth of the second chamber portion 22 were 1.1 mm or more, the thickness of the body portion 10 would need to be further reinforced, which would reduce the heat transfer efficiency to the reaction reagent. According to this embodiment, the depth of the second chamber portion 22 is 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, multiple reaction chambers 20 are provided, and different reaction reagents are immobilized in the reaction chambers 20. This allows for the detection of different target genes through a single sample injection, enabling multiple diagnoses. For example, some of the multiple second chambers 22 may contain and immobilize a reaction reagent for detecting a specific target gene. Other second chambers 22 may contain and immobilize reaction reagents for detecting other target genes. In this case, each reaction reagent may be immobilized to contain a fluorescent dye. It goes without saying that the fluorescent dye may be injected into the chip together with the sample. The fluorescent dye immobilized in each of the different reaction reagents or injected together with the sample may be the same dye. When the fluorescent dye reacts with the target gene in each second chamber 22 and amplifies it, the light emitted is detected, allowing the reaction in that second chamber 22 to be identified. In other words, multiple diagnoses can be performed at a time by irradiating a single wavelength.

[0051] The reaction reagent is immobilized in the second chamber 22, and the chip with the immobilized reaction reagent can be transported to a location where a PCR test is required and used as needed. The reaction reagent can be immobilized in the second chamber 22 by methods such as air drying, vacuum drying, or freeze drying. The surface of the reaction reagent immobilized in the second chamber 22 can be coated with a waterproof and adhesive coating agent. The coating agent can be immobilized with the reagent (primers, probes, premix) using a biopolymer that does not affect the reagent properties or PCR results. The coating agent can remain solid at room temperature and melt above a certain temperature. For example, the coating agent can melt above 45°C. During PCR testing, when a chip with a sample injected therein is heated, the coating agent melts, allowing the sample and the reaction reagent to react. 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 unit 22. Even in such a case, the reaction reagent accommodated in the second chamber unit 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 melts and the sample and the reaction reagent may react.

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

[0054] 4, the first inclined channel 221 is formed so that its width increases and its shape becomes curved as it approaches the inner circumferential surface of the second chamber section 22. That is, the left and right side surfaces of the first inclined channel 221 are formed so that their width increases and their shape becomes curved in the direction of sample inflow near the portion where the first inclined channel 22 is connected to the second chamber section 22. Similarly to the first inclined channel 221, the second inclined channel 222 is formed so that its width increases and its shape becomes curved as it approaches the inner circumferential surface of the second chamber section 22. The second inclined channel 222 is disposed opposite the first inclined channel 221, and its left and right side surfaces are formed so that their width increases and their shape becomes curved as they approach the inner circumferential surface of the second chamber section 22.

[0055] The inlet channel 30 connects the inlet 11 and the reaction chamber 20 and guides the sample to the reaction chamber 20. According to the present embodiment, the inlet 11 protrudes 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 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 channel with the outside air when the sample is injected through the inlet 11, allowing the sample to flow along the channel 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 one another by connecting channels 40 so that the sample flows in one direction.

[0057] According to this embodiment, as shown in FIGS. 2 and 3, the reaction chambers 20 include a first row and a second row. In the first row, the reaction chambers 20 are spaced apart from one another in a first direction. The reaction chambers 20 in the first row are arranged on an imaginary 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 in the second row are also arranged on an imaginary line extending in one direction, similar to the first row, and the imaginary lines on which the first and second rows are arranged are parallel to each other. With this structure, the reaction chambers 20 in the first row and the reaction chambers 20 in the second row are arranged in a zigzag pattern. According to this embodiment, the first column includes seven reaction chambers 20, and the second column includes six reaction chambers 20, forming a total of 13 reaction chambers 20. As described above, the same or different reaction reagents can be placed in each reaction chamber 20. According to this embodiment, up to 13 reaction reagents can be immobilized in the reaction chambers 20, and multiple target genes can be distinguished and multiple diagnoses can be performed 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 plurality of 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 proximity to the inlet 11, the connecting channel 40 connects the N-th chamber to the (N+1)-th chamber. Here, "in order of proximity to the inlet 11" refers to the order in which the sample passes when it passes through the reaction chambers 20 sequentially via the connecting channel 40, starting from the reaction chamber 20 to which it first reaches via the inlet 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] One end of the first flow path 41 is connected to the Nth chamber, and the other end extends toward the (N+1)th chamber. The other end of the second flow path 42 is connected to the (N+1)th chamber, and one end extends 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 this 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 approximately 90 degrees, and the second bending portion 4312 changes the flow direction of the sample by 180 degrees. According to this embodiment, the sample flowing out of the Nth chamber changes direction by approximately 90 degrees through the first bending portion 4311, then passes through the second bending portion 4312 three times in succession, and then changes direction by approximately 90 degrees through the first bending portion 4311 again to be introduced into the (N+1)th chamber. According to this embodiment, the bending portion 431 is formed with two first bending portions 4311 and three second bending portions 4312, but the number of first bending portions 4311 and second bending portions 4312 is not limited thereto and may be formed in various ways.

[0062] The bent portion 431 prevents the sample from moving between adjacent reaction chambers 20 when the sample is injected into each reaction chamber 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 mixing 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 the inlet side where the 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 as the sample flows into the reaction chamber 20, it slows down along the section where the width increases and fills the interior 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 as 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, increasing the flow rate of the sample and facilitating the transfer of the sample to other adjacent reaction chambers 20.

[0066] According to this embodiment, the depth of the connecting channel 40 recessed from the bottom surface of the body portion 10 is 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 deeper than the connecting channel 40, and the step between the first chamber portion 21 and the second chamber portion 22 is formed to the same depth as the connecting channel 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 inlet channel 30, the connecting channel 40, and the reaction chamber 20, which are groove-shaped and formed from the bottom surface of the body part 10 to the top surface. The cover part 50 is attached to the bottom surface of the body part 10 with the sample accommodated and fixed in the reaction chamber 20. The cover part 50 forms the bottom surfaces of the inlet 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 via the inlet channel 30 and the connecting channel 40, and at this time, the bottom surfaces of the inlet 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 50 is disposed under the body 10 and is provided in the form of a film so that heat can be transferred from the heating device 120 for PCR. The cover 50 is made of a thin film, which can easily transfer heat from the heating device 120 and can be made of a metal or a polymeric material containing nanocarbon, which has excellent heat transfer efficiency. The sample moves automatically through the inlet channel 30, the connecting channel 40, and the reaction chamber 20 without using any power.

[0070] The actions and effects 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 underside of the body 10 according to an embodiment of the present invention. First, a reaction reagent is immobilized in the second chamber 22 of the reaction chamber 20. Different reaction reagents for detecting different target genes may be immobilized in the reaction chambers 20. The underside of the body 10 is sealed by a film-type cover 50. The cover 50 closes the inlet channel 30, the connecting channel 40, and the reaction chamber 20. According to this embodiment, adhesive regions are provided on the edges of the inlet channel 30, the connecting channel 40, and the reaction chamber 20 so that the film-type cover 50 can be attached. The adhesive regions are wider than the inlet channel 30 and the connecting channel 40 to prevent sample leakage during sample injection. The adhesive regions may be formed along the edges of the inlet channel 30, the connecting channel 40, and the reaction chamber 20, with a width of 0.8 mm to 1.0 mm. If the width of the adhesive area 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 immobilized in the reaction chamber 20 together with a coating agent. The chip on which the reaction reagent is immobilized 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 using a PCR device. Therefore, the PCR test chip 100 according to the present invention allows reaction reagents to be immobilized in advance, allowing PCR tests to be performed quickly, and allows multiple reaction reagents to be immobilized in advance, allowing multiple tests to be performed.

[0073] A sample is injected through the inlet 11. Two protrusions are formed on the top surface of the body 10, either of which 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 10. The remaining inlet 11 is connected to the outside air, and the sample itself moves along a narrow channel.

[0074] The sample is introduced into the reaction chamber 20 along the inlet channel 30, and then into the successively arranged reaction chambers 20. From the Nth chamber, the sample flows out along the first channel 41. As the sample leaves the Nth chamber, its movement speed increases as it passes through the second expansion section 24, allowing it to easily move to the (N+1)th chamber. The sample then changes its flow path as it passes through the bent section 431 of the direction-changing channel 43. The bent section 431 is located between the Nth chamber and the (N+1)th chamber, forming a pressure boundary that prevents the sample from moving to adjacent chambers if it reacts with a reaction reagent. After passing through the direction-changing channel 43, the sample is introduced into the (N+1)th chamber via the second channel 42. According to this embodiment, the sample flows from the first chamber section 21 to the thirteenth chamber along the path described above.

[0075] Furthermore, when a sample is introduced into the Nth chamber, the sample flows into the inlet and also flows along the first inclined channel 221. The first inclined channel 221 is formed so that it becomes deeper toward the second chamber portion 22, the width between the side portions becomes wider, and it 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 this embodiment is formed in two stages by a first chamber portion and a second chamber portion. Since the reaction reagent is fixed in the second chamber portion 22, bubbles are generated as the reaction reagent comes into contact with the sample and expands, and the bubbles are collected in the second chamber portion 22. The sample entering the second chamber portion 22 along the first inclined channel 221 can push the bubbles toward the second inclined channel 222 and remove them. Since the inner surface of the second chamber portion 22 is circular, the sample moves along the streamlined inner surface, effectively removing the bubbles. By removing the bubbles, the reaction result between the sample and the reaction reagent can be accurately obtained. The first inclined channel 221 and the second inclined channel 222 are also provided to remove PCR inhibitors, such as bubbles, that may be 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, which allows the fluorescent material to be concentrated in a limited space, thereby improving the sensitivity of the reaction results during image acquisition and analysis through a PCR device and improving diagnostic accuracy. In addition, because the reaction reagent reacts with the sample while being fixed in the well in the second chamber 22, movement of the reaction sample between the reaction chambers 20 is suppressed, and the reaction samples that react with the sample do not mix with each other, further improving the reliability of the reaction results.

[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 carrying out PCR by mounting the aforementioned 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 for providing heat to the chip 100, a light source 130 for irradiating light onto the chip 100, a camera 140 for capturing fluorescence generated when the reaction reagent and the sample react to obtain an image, and an analysis unit 150 for analyzing the image of the camera 140.

[0080] The main body 110 includes a mounting part to 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 plurality 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 on the underside of the chip 100 and heats the reaction reagents and samples to a temperature required for PCR testing. At this time, the heat from the heating means is transferred to the reaction reagents and samples via the film-type cover part 50, allowing for rapid heating.

[0082] The light source 130 is provided to irradiate light onto 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 have a single wavelength or different wavelengths.

[0083] The camera 140 is provided to capture the fluorescence emitted by the fluorescent dye in response to 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 once. The camera 140 can capture fluorescent images at regular intervals above the chip 100.

[0084] The analysis unit 150 analyzes the image acquired by the camera 140. The analysis unit 150 compares the hue of the image with predetermined reference data for analysis. The predetermined reference data is data in which various target genes or diseases are pre-matched according to the type of color (RGB ratio) and intensity of emitted fluorescent light. The analysis unit 150 can calculate the RGB ratio of the image acquired 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 this process, the present invention enables multiplex testing by detecting various types of target genes with a single 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 by citing preferred embodiments, but the present invention is not limited to the above embodiments, and various modifications can be provided within 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 introduced; a reaction chamber including a first chamber portion recessed from the lower surface of the body portion to the 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 in the first region, for accommodating a reaction reagent; an inlet channel connecting the inlet and the reaction chamber and guiding the sample to the reaction chamber; a plurality of reaction chambers are provided in the body portion; The reaction chambers are connected to each other by a connecting channel so that the sample flows in one direction; 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; a direction-changing flow channel that connects the first flow channel and the second flow channel and has a bent portion that changes the flow direction of the sample.

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

3. The reaction chamber comprises: the reaction chambers are arranged to form a first row in which they 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 arranged 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.

4. 2. The PCR testing chip according to claim 1, wherein the number of the bent portions is at least two.

5. a plurality of reaction chambers are provided 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 portion that closes 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 portion to the upper surface of the body portion, The PCR testing chip according to claim 1 , wherein the cover forms the bottom surfaces of the inlet channel, the connecting channel, and the reaction chamber.

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

7. A body portion made of a light-transmitting material and having an inlet through which a sample is introduced; a reaction chamber including a first chamber portion recessed from the lower surface of the body portion to the 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 in the first region, for accommodating a reaction reagent; an inlet channel connecting the inlet and the reaction chamber and guiding the sample to the reaction chamber; a plurality of reaction chambers are provided 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 portion that closes 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 portion to the upper surface of the body portion, the cover part forms the inlet channel, the connecting channel, and a bottom surface of the reaction chamber; The PCR testing chip according to claim 1, wherein the depth of the connecting channel recessed from the lower surface of the body portion is equal to the depth of the first chamber portion.

8. the second chamber portion includes a first inclined flow channel formed in a direction in which the sample is introduced, and a second inclined flow channel formed in a direction in which the sample is discharged from the second chamber portion; the first inclined flow passage is formed such that its depth from the lower 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 it approaches the inner peripheral surface, 2. The PCR testing chip according to claim 1, wherein the second inclined flow channel is formed opposite to the first inclined flow channel, and is formed such that the depth from the bottom surface of the body portion gradually increases as the second inclined flow channel 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.

9. 9. The PCR testing chip according to claim 8, 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 that they are curved.

10. the connecting channel has a first expansion part at an inlet side through which the sample is introduced into the reaction chamber, the first expansion part having a width that gradually increases as the width approaches the reaction chamber; 2. The PCR testing chip according to claim 1, wherein the connecting channel has a second expansion part at a side of an outlet through which the sample is discharged from the reaction chamber, the second expansion part having a width that gradually increases as the connecting channel approaches the reaction chamber.

11. the reaction reagents include at least two or more types of reaction reagents so as to detect different target genes, 3. The PCR testing chip according to claim 2, wherein different kinds of reaction reagents are accommodated in the plurality of reaction chambers to enable a plurality of diagnoses for one sample.

12. a main body; 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 plurality of reaction chambers, each having a first chamber portion recessed from the lower surface of the body portion toward the upper surface of the body portion in a first region of the lower surface of the body portion from the lower surface toward the upper surface of the body portion, and a second chamber portion in which a reaction reagent is accommodated and further recessed from the first chamber portion toward the upper surface of the body portion in the first region; an inflow channel connecting the inlet and the reaction chambers and guiding the sample to the reaction chambers; and a connecting 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 portion and 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, 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; a direction-changing flow channel that connects the first flow channel and the second flow channel and has a bent portion that changes the flow direction of the sample.

13. The reaction chamber comprises: the reaction chambers are arranged to form a first row in which they 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 arranged between the chambers in the first row; 13. The PCR testing device according to claim 12, wherein the reaction chambers forming the first row and the reaction chambers forming the second row are arranged in a zigzag pattern.

14. a cover attached to the lower surface of the body portion and formed in a groove shape from the lower surface of the body portion to the upper surface of the body portion, the inlet channel, the connecting channel, and the reaction chamber; the cover portion is formed in a film shape 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 12, wherein the cover forms the bottom of the inlet channel, the connecting channel, and the reaction chamber.

15. the second chamber portion includes a first inclined flow channel formed in a direction in which the sample is introduced, and a second inclined flow channel formed in a direction in which the sample is discharged from the second chamber portion; the first inclined flow passage is formed such that its depth from the lower 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 it approaches the inner peripheral surface, 13. The PCR testing device according to claim 12, wherein the second inclined flow path is formed opposite to the first inclined flow path, and is formed such that the depth from the bottom surface of the body portion gradually increases from a point spaced a predetermined distance from the second chamber portion to the inner surface of the second chamber portion as the distance approaches the inner surface.

16. the connecting channel has a first expansion part at an inlet side through which the sample is introduced into the reaction chamber, the first expansion part having a width that gradually increases as the width approaches the reaction chamber; 13. The PCR testing device of claim 12, wherein the connecting channel has a second expansion portion at a side of an outlet through which the sample is discharged from the reaction chamber, the second expansion portion having a width that gradually increases as the connecting channel approaches the reaction chamber.

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