Multi-functional PCR test chip

Abstract

The present disclosure is characterized by including a body portion including a light-transmissive material, a sample inlet through which a sample is introduced, and a processing-solution inlet through which a processing solution for processing the sample is introduced, a sample test chamber portion provided in the body portion and including a reaction chamber, the reaction chamber accommodating a reaction reagent for detecting a target genetic material and in which the sample introduced through the sample inlet reacts with the reaction reagent, and a processing-solution test chamber portion provided in the body portion and including a test chamber, the test chamber accommodating the reaction reagent and in which the processing solution introduced through the processing-solution inlet reacts with the reaction reagent.

Description

Technical Field

[0001] The present disclosure relates to a multi-functional PCR test chip for reacting a sample with a reaction reagent to perform a PCR test. The reaction reagent is fixed to the chip, the sample to be tested is introduced into the chip, and a PCR test is performed by checking whether the sample reacts with the reaction agent.Background Art

[0002] Polymerase chain reaction (hereinafter referred to as PCR) refers to a method for amplifying a specific target genetic material to be detected, that is, a technique for amplifying a small amount of genetic material to obtain a large amount of genetic material having the same base sequence as the original genetic material. PCR is used to amplify human nucleic acids to diagnose various types of genetic diseases or is applied to nucleic acids of bacteria, viruses, or fungi to diagnose infectious diseases.

[0003] In general, PCR is performed by repeatedly carrying out the following three steps. The three steps include: 1) a denaturing step of heating a sample solution containing double-stranded DNA to a specific temperature, for example, about 95 °C, to separate the double-stranded DNA into single-stranded DNA; 2) an annealing step of, after the denaturing step, providing the sample solution with an oligonucleotide primer having a sequence complementary to a specific base sequence to be amplified, and cooling the sample solution together with the single-stranded DNA to a specific temperature, for example, 55 °C, to bind the primer to the specific base sequence of the single-stranded DNA to form a partial DNA-primer complex; and 3) an extension (or amplification) step of, after the annealing step, maintaining the sample solution at an activation temperature of a DNA polymerase, for example, 72 °C, to form double-stranded DNA using the DNA polymerase based on the primer of the partial DNA-primer complex. A target nucleic acid having the specific base sequence may be exponentially amplified by repeating these three steps multiple times. However, in the case of RNA, a step of reverse-transcribing the RNA into DNA may be performed prior to the steps 1) to 3).

[0004] To perform such PCR, a PCR chip equipped with a chamber capable of accommodating a reagent and a sample and a PCR device configured to heat and cool the PCR chip to induce an amplification reaction and to measure a result thereof are required. As devices for amplifying target nucleic acids, various PCR devices have been developed.

[0005] In the related art, after completion of PCR, only qualitative analysis of amplified target nucleic acids through a separate electrophoresis process was possible. However, real-time PCR devices (that is, rPCR devices) have recently been developed, which enable quantitative analysis of target nucleic acids by detecting fluorescence intensity proportional to the concentration of amplified genetic material using an optical detection system. In addition, portable real-time PCR devices have been developed by miniaturizing large PCR devices used in laboratories, thereby enabling PCR at a sample collection site. Portable PCR devices are characterized by the use of PCR chips (or biochips or micro-chips) and cameras or digital cameras instead of large and complex optical detection systems.

[0006] The present disclosure relates to a PCR test chip that enhances sensitivity when detecting the intensity of fluorescence emitted during a reaction between a reagent and a sample, enables simultaneous detection of multiple target genes, and enables simultaneous checking for contamination of a processing solution such as a buffer used for extracting nucleic acids from a sample.Disclosure of Invention Technical Problem

[0007] In the related art, a processing solution (buffer solution) is used to extract nucleic acids from a sample, and nucleic acids included in the buffer solution are introduced into a chip to test whether a reaction with reaction reagents occurs. However, there is an inconvenience in that contamination of the processing solution is not separately checked, and an additional procedure is performed to verify such contamination.

[0008] The present disclosure is provided to address the above-described problems. An objective of the present disclosure is to provide a PCR test chip in which reaction reagents that react with a sample are fixed to reaction chambers, thereby enhancing sensitivity in checking test results and enabling contamination of a processing solution, such as a buffer used to extract nucleic acids from a sample, to be checked together.Solution to Problem

[0009] According to an embodiment of the present disclosure, a multi-functional PCR test chip includes: a body portion including a light-transmissive material, a sample inlet through which a sample is introduced, and a processing-solution inlet through which a processing solution for processing the sample is introduced; a sample test chamber portion provided in the body portion and including a reaction chamber, the reaction chamber accommodating a reaction reagent for detecting a target genetic material and in which the sample introduced through the sample inlet reacts with the reaction reagent; and a processing-solution test chamber portion provided in the body portion and including a test chamber, the test chamber accommodating the reaction reagent and in which the processing solution introduced through the processing-solution inlet reacts with the reaction reagent.

[0010] In addition, the reaction chamber may preferably include: an inlet portion through which the sample flows in; a reaction well portion in which the sample reacts with the reaction reagent, the reaction well portion being recessed upward from a lower surface of the body portion; and an outlet portion through which the sample flows out from the reaction well portion.

[0011] In addition, preferably, the inlet portion and the outlet portion of the reaction chamber may be inclined.

[0012] In addition, preferably, a plurality of reaction chambers may be provided in the body portion, and the plurality of reaction chambers may be connected to each other through a sample flow path to allow the sample to flow in one direction.

[0013] In addition, preferably, the inlet portion may include an expanded portion that widens in a direction in which the sample enters the reaction well portion, and the outlet portion may include a reduced portion that narrows in a direction in which the sample flows out from the reaction well portion.

[0014] In addition, preferably, when the plurality of reaction chambers are referred to as N-th chambers (where N is a natural number) in order adjacent to the sample inlet, the sample flow path may connect an N-th chamber and an (N+1)-th chamber to each other, wherein the sample flow path may include: a first flow path having one end connected to the N-th chamber; a second flow path having another end connected to the (N+1)-th chamber; and a first connection flow path provided between the first flow path and the second flow path and connecting the first flow path and the second flow path to each other, wherein the first flow path and the second flow path may include at least one bending portion to change a flow direction of the sample.

[0015] In addition, preferably, a cover portion may be coupled to a lower surface of the body portion to close the sample test chamber portion and the processing-solution test chamber portion that are provided as grooves recessed upward from the lower surface of the body portion, and in a state in which the body portion is positioned with the cover portion facing downward, the cover portion may form bottom surfaces of the sample test chamber portion and the processing-solution test chamber portion.

[0016] In addition, preferably, the cover portion may be provided as a film disposed on a lower side of the body portion and attached to the lower surface of the body portion to transfer heat from a heating device for performing PCR.

[0017] In addition, preferably, the test chamber may include a plurality of test chambers.

[0018] In addition, preferably, the plurality of test chambers may be connected to each other through a processing-solution flow path to allow the processing solution to flow in one direction.

[0019] In addition, preferably, when the plurality of test chambers are referred to as M-th chambers (where M is a natural number) in order adjacent to the processing-solution inlet, the processing-solution flow path may connect an M-th chamber and an (M+1)-th chamber to each other, wherein the processing-solution flow path may include: a third flow path having one end connected to the M-th chamber; a fourth flow path having another end connected to the (M+1)-th chamber; and a second connection flow path provided between the third flow path and the fourth flow path and connecting the third flow path and the fourth flow path to each other, wherein the third flow path and the fourth flow path may include at least one bending portion to change a flow direction of the sample.

[0020] In addition, preferably, the plurality of reaction chambers may be arranged adjacent to one another, and the test chamber may include a plurality of test chambers, at least one of the plurality of test chambers being provided on each of both outermost sides of the plurality of reaction chambers arranged adjacent to one another.

[0021] In addition, preferably, the sample test chamber portion may include: a first sample test chamber portion including a plurality of reaction chambers arranged adjacent to each other; and a second sample test chamber portion provided apart from the first sample test chamber portion and including another plurality of reaction chambers arranged adjacent to each other, wherein the processing-solution test chamber portion may include at least one first test chamber adjacent to the first sample test chamber portion and at least one second test chamber adjacent to the second sample test chamber portion.

[0022] In addition, preferably, the first sample test chamber portion and the second sample test chamber portion may be arranged adjacent to each other between the first test chamber and the second test chamber.

[0023] In addition, preferably, reaction reagents for detecting at least two different target genes may be fixed to the reaction chamber and the test chamber to enable multiplex diagnosis for samples.Advantageous Effects of Invention

[0024] An embodiment of the present disclosure provides a PCR test chip in which reaction reagents that react with a sample are fixed to reaction chambers, thereby improving sensitivity in checking test results and enabling contamination of a processing solution, such as a buffer used to extract nucleic acids from a sample, to be checked together.

[0025] Specifically, the present disclosure aims to enable simultaneous multiplex diagnosis with high sensitivity and specificity. In multiplex PCR diagnosis of the related art, a plurality of primer probes for detecting multiple targets are disposed in the same space, leading to interference, competitive reactions, and variations in reaction characteristics, thereby degrading sensitivity and specificity. To address this problem, target detection regions are spatially separated from each other to eliminate mutual interference and competitive reactions.

[0026] In addition, unlike multiplex PCR diagnosis of the related art, there is no need for multiple preparations of reaction solutions, and because the reaction solutions are connected to each other through a single flow path, a reaction may be performed simply and rapidly with a single preparation and injection of reaction solutions.

[0027] Further, although the reaction solutions are connected to each other through a single flow path, diffusion or interference between target spaces are prevented by microfluidic techniques, thereby improving the accuracy of test results.

[0028] Further, a flow path for contamination checking (NC: negative control) and a flow path for verification of equipment and reagents (PC: positive control) are separately designed, thereby providing an effect of enabling not only inspection of a sample but also NC testing and / or PC testing to be performed together.

[0029] Further, a sample flow path, through which reaction chambers are connected to each other, and a processing-solution flow path, through which test chambers are connected to each other, are provided with bending portions, thereby effectively preventing a processing solution or a sample from moving from one reaction chamber or test chamber to an adjacent reaction chamber or test chamber when the processing solution or sample reacts with reaction reagents. As a result, the accuracy of test results may be improved.

[0030] Further, because the reaction chambers or test chambers include recessed wells for reaction reagents, reaction may occur in the wells, thereby improving visibility. In addition, each of the reaction chambers or test chambers is provided with an expanded portion at an inlet and a reduced portion at an outlet, and thus, a sample or processing solution may be rapidly dispensed (diffused).Brief Description of Drawings

[0031] FIG. 1 is a perspective view illustrating a multi-functional PCR test chip according to an embodiment of the present disclosure, FIG. 2 is a bottom perspective view of FIG. 1, FIG. 3 is an enlarged view of FIG. 2, FIG. 4 is a view illustrating functions of a reaction chamber and a test chamber of FIG. 1, FIG. 5 is a cross-sectional view of FIG. 1, FIG. 6 is a perspective view illustrating a multi-functional PCR test chip according to another embodiment of the present disclosure, FIG. 7 is a bottom perspective view of FIG. 6, FIG. 8 is a view illustrating functions of a reaction chamber and a test chamber of FIG. 7, and FIG. 9 is a view illustrating a multi-functional PCR test chip according to still another embodiment of the present disclosure. Mode for the Invention

[0032] Hereinafter, various embodiments of the present disclosure will be described with reference to the accompanying drawings. Various embodiments of the present disclosure may be subject to various modifications and may have various forms, and specific embodiments are illustrated in the drawings and described in detail. However, this is not intended to limit the various embodiments of the present disclosure to specific forms, and all modifications and / or equivalents or substitutes within the spirit and technical scope of the various embodiments should be considered as being included in the scope of the present disclosure. In the drawings, like reference numerals denote like elements.

[0033] In various embodiments of the present disclosure, expressions such as "include" or "may include" specify the presence of the disclosed corresponding functions, operations, or elements, and do not preclude the presence of one or more additional functions, operations, or elements. Furthermore, in various embodiments of the present disclosure, terms such as "include" or "have" are intended to specify the presence of features, numbers, steps, operations, elements, components, or combinations thereof described in the specification, and do not to preclude the presence or addition of one or more other features, numbers, steps, operations, elements, components, or combinations thereof.

[0034] It should be understood that when an element is referred to as being "connected to" another element, the element may be directly connected to the other element, or any other element may be interposed between the two elements. In contrast, it should be understood that when an element is referred to as being "directly connected to" or "directly coupled to" another element, there is no other element between the two elements.

[0035] Terms used in various embodiments of the present disclosure are only for describing specific embodiments and are not intended to limit the various embodiments of the present disclosure. The terms of a singular form may include plural forms unless referred to the contrary.

[0036] Unless defined otherwise, all terms used herein, including technical and scientific terms, have the same meaning as those commonly understood by a person skilled in the art to which various embodiments of the present disclosure pertain.

[0037] Terms defined in generally used dictionaries should be interpreted as having meanings consistent with the context of the related art, and unless explicitly defined in various embodiments of the present disclosure, should not be interpreted as having idealized or excessively formal meanings.

[0038] The present disclosure relates to a chip used in a PCR device. The chip accommodates a reaction reagent, and after a sample is injected into the chip, a target gene is detected using the PCR device. The present disclosure provides a structure in which a portion having a plurality of reaction chambers into which a sample is injected is separated from a chamber portion for testing contamination of an introduced processing solution, a reaction reagent, or the like to enable negative control (NC) testing and / or positive control (PC) testing, thereby enabling simultaneous inspection of the sample and the processing solution.

[0039] In the present specification, the term "polymerase chain reaction (PCR)" refers to a reaction for amplifying a specific target nucleic acid molecule by using a thermostable DNA polymerase. For PCR, a reaction mixture including, in addition to a DNA polymerase, primers (forward primers and reverse primers), which are oligonucleotides capable of hybridizing specifically with a target nucleic acid, a deoxynucleotide triphosphate (dNTP) mixture, and divalent ions such as Mg 2+< may be used.

[0040] A "primer" is used to initiate a PCR process and refers to an oligonucleotide or polynucleotide that hybridizes complementarily with a template DNA. As primers for a PCR process, a forward primer (or sense primer) selected from a sense strand having the same direction as the genetic code progression of a nucleic acid molecule to be amplified, and a reverse primer (or antisense primer) selected from an antisense strand complementary to the sense strand, may be used as a pair.

[0041] The term "sample" refers to a genetic material to be amplified or a biological solution containing such genetic material. The term "reaction reagent" refers to a reagent that is used to detect a target genetic material and may include primers or the like. Primers may include a pair of primers having a length of 15 bp to 30 bp and capable of binding to both ends of a specific region of a target gene. In addition, an enzyme that does not lose activity even at a high temperature of 90 °C or higher is used as a DNA polymerase.

[0042] Hereinafter, preferred embodiments of the present disclosure will be described in detail with reference to the accompanying drawings. According to an embodiment of the present disclosure, a multi-functional PCR test chip 1001 includes a body portion 100, a sample test chamber portion 200, and a processing-solution test chamber portion 300.

[0043] The body portion 100 may include a light-transmissive material to detect a fluorescent substance by using an optical detection device. As illustrated in FIG. 1, according to the present embodiment, the body portion 100 may include a transparent polymer material. The body portion 100 may have a rectangular shape. The body portion 100 may have various shapes such as a circular shape or a square shape. According to the present embodiment, protruding walls 13 are formed on opposite long sides of the rectangular shape of the body portion 100 such that a user may easily grip the body portion 100. Reinforcing protrusions 14 extending in a vertical direction are formed on inner sides of the protruding walls 13. The reinforcing protrusions 14 prevent an injected material from bending when the body portion 100 is formed by injection molding, and improve the rigidity of the body portion 100. In addition, during product manufacture or in the field, the reinforcing protrusions 14 allow chips to be stacked in a vertical direction, and in this case, the reinforcing protrusions 14 may function as supports for supporting an upper chip. In addition, grooves are formed in outer peripheral surfaces of the protruding walls 13 such that a user may easily grip the body portion 100.

[0044] In addition, as illustrated in FIGS. 1 and 2, the body portion 100 includes sample inlets 11 into which a sample is introduced, and processing-solution inlets 12 into which a processing solution for processing the sample is introduced. Flow paths 40 and 50 and a plurality of chambers 20 and 30, all of which are shaped like grooves, are formed in the body portion 100. A sealing film is attached to the body portion 100, and together with the flow paths 40 and 50 and the chambers 20 and 30, the sealing film forms flow paths through which a sample and a processing solution move.

[0045] The sample test chamber portion 200, which is provided in the body portion 100, includes the reaction chamber 20 in which a reaction reagent for detecting a target genetic material is accommodated such that a sample introduced through the sample inlets 11 may react with the reaction reagent. According to the present embodiment, the sample test chamber portion 200 may include a flow path extending in one direction and a plurality of reaction chambers 20 provided along the flow path.

[0046] As illustrated in FIG. 2, each of the reaction chambers 20 includes an inlet portion 21 through which a sample flows in, a reaction well portion 22 recessed upward from a lower surface of the body portion 100 to allow a reaction between the sample and a reaction reagent, and an outlet portion 23 through which the sample flows out from the reaction well portion 22. Each of the reaction chambers 20 may include a reaction reagent that reacts with a sample. Specifically, the reaction reagent that reacts with the sample may be fixed to the reaction well portion 22.

[0047] According to the present embodiment, a plurality of reaction chambers 20 are provided in the body portion 100. The reaction chambers 20 are connected to each other through a sample flow path 40 to allow a sample to flow in one direction. That is, the sample flow path 40 and the reaction chambers 20 are connected in series to each other. The sample inlets 11, which are provided to receive a sample, are connected to both ends of the reaction chambers 20 that are connected in series to each other. When testing a sample, a user may inject the sample through any one of the sample inlets 11.

[0048] The reaction reagent may include a primer or the like. In addition, the reaction reagent may further include a fluorescent dye. Depending on a PCR method, the fluorescent dye may be injected into the chip together with a sample. The fluorescent dye emits light in a specific wavelength band during a DNA extension (or amplification) step. At least two types of reaction reagents may be provided to detect different target genes, and different reaction reagents may be accommodated in the reaction chambers 20. Because a plurality of reaction chambers 20 are provided, different reaction reagents may be fixed to the reaction chambers 20 to enable detection of different target genes with a single injection of a sample.

[0049] For example, in an embodiment, six reaction chambers 20 may be provided. When reaction reagents that each detect two types of specific target genes are fixed to the reaction chambers 20, respectively, a total of twelve target genes may be detected. In this case, the reaction reagents may be fixed to the reaction chambers 20 in a state in which each of the reaction reagents includes a fluorescent dye. Alternatively, the fluorescent dye may be injected into the chip together with a sample. Fluorescent dyes may be provided by fixing a fluorescent die together with different reaction reagents, injecting a first die together with a sample, or injecting different first and second dies together with a sample. The fluorescent dyes respond to light of a specific wavelength, and the first and second dyes may generate fluorescence with respect to light of different wavelengths. When amplification is performed by a reaction with a target gene in each the reaction chambers 20, a reaction in the reaction chamber 20 may be identified by detecting emission of light. When the first dye is injected identically into six chambers 20, six types of multiplex diagnosis may be possible at once by projecting light of a single wavelength to which the first dye responds. When the first dye and the second dye are simultaneously injected into each of the reaction chambers 20, twelve types of multiplex diagnosis may be possible at once by projecting multi-wavelength light having two different wavelengths.

[0050] The reaction reagents are fixed to the reaction chambers 20, and the chip in which the reaction reagents are fixed may be transported to a place at which PCR testing is required and used as needed. The reaction reagents may be fixed to the reaction chambers 20 by a method such as natural drying, vacuum drying, or freeze-drying. In addition, surfaces of the reaction reagents fixed to the reaction chambers 20 may be coated with a waterproof and adhesive coating agent.

[0051] The coating agent may be fixed together with reagents (primers, probes, or premixes) by using a biopolymer material that does not affect properties of the reagents or PCR results. The coating agent may maintain a solid state at room temperature and may melt at a temperature equal to or higher than a predetermined temperature. For example, the coating agent may melt at 45 °C or higher. When the chip into which a sample is injected is heated during a PCR process, the coating agent may melt, allowing a reaction between the sample and a reaction reagent. The coating agent may include any one of wax, agarose, paraffin, collagen, chitosan, and gelatin, or a mixture thereof. The coating agent may be fixed to a reaction chamber in a state in which the coating agent is mixed with a reaction reagent. Even in this case, the reaction reagent accommodated in the reaction chamber may maintain a solid state at room temperature, and when the chip into which the sample is injected is heated during a PCR process, the coating agent may melt to allow a reaction between the sample and the reaction reagent.

[0052] According to the present embodiment, the sample flow path 40, through which a sample is transported, connects the reaction chambers 20 to each other and includes at least one curved portion in a section connecting adjacent reaction chambers 20. The sample flow path 40 preferably has a size of less than 0.55 mm to prevent excessive pressure during movement of a sample and thus ensure smooth movement thereof. Each of the reaction chambers 20 is provided in the form of a groove extending in one direction.

[0053] The inlet portion 21 of each of the reaction chambers 20 includes an expanded portion 211 that gradually widens in a direction toward the reaction chamber 20. The expanded portion 211 is tapered such that when a sample flows into the reaction chamber 20, the sample is decelerated by a widening section and fills the internal space of the reaction chamber 20. The outlet portion 23 of each of the reaction chambers 20 includes a reduced portion 231 that gradually narrows in a direction away from the reaction chamber 20 for a sample flowing out from the reaction chamber 20. Owing to the reduced portion 231, the flow speed of a sample may increase, and thus, the sample may easily flow to an adjacent reaction chamber 20.

[0054] A bottom surface of the reaction chamber 20 is deeper than a bottom surface of the sample flow path 40. The inlet portion 21 includes an inclined surface 24 connected to the bottom surface of the reaction chamber 20, and the outlet portion 23 similarly includes an inclined surface 24 connected to the bottom surface of the reaction chamber 20. The bottom surface of the reaction chamber 20 is deeper than the bottom surface of the sample flow path 40, thereby forming a sufficient space for fixing a reaction reagent, and because the reaction reagent is fixed to the deeper bottom surface, the reaction agent may have improved reactivity with the sample. When a fluorescent substance is used, the fluorescent substance may be more easily checked because the reaction well portion 22 is deepened.

[0055] According to the present embodiment, the sample flow path 40 is provided to connect a plurality of reaction chambers 20 to each other. When the reaction chambers 20 are referred to as N-th chambers (where N is a natural number) in an order adjacent to a sample inlet 11, the sample flow path 40 connects an N-th chamber and an (N+1)-th chamber. Here, the expression "in the order adjacent to a sample inlet 11" refers to an order in which a sample passes through the reaction chambers 20 sequentially through the sample flow path 40, starting from a reaction chamber 20 that the sample first reaches after being injected through the sample inlet 11. According to the present embodiment, the sample flow path 40 includes a first flow path 41, a second flow path 42, and a first connection flow path 43.

[0056] The first flow path 41 has one end connected to the N-th chamber and the other end extending toward the (N+1)-th chamber. The second flow path 42 has one end connected to the (N+1)-th chamber and the other end extending toward the N-th chamber. The first connection flow path 43 is provided between the first and second flow paths 41 and 42 to connect the first and second flow paths 41 and 42 to each other. That is, one end of the first connection flow path 43 is connected to the other end of the first flow path 41, and the other end of the first connection flow path 43 is connected to the end of the second flow path 42. According to the present embodiment, the first and second flow paths 41 and 42 include a bending portion 44 for changing the flow direction of a sample. At least one bending portion 44 may be provided.

[0057] According to the present embodiment, as illustrated in FIG. 3, the sample flow path 40 is designed such that a sample changes direction three times while moving from the first flow path 41 to the first connection flow path 43, and three times while moving from the first connection flow path 43 to the second flow path 42. However, the number of bending portions 44 is not limited thereto and may vary. The bending portions 44 prevent a sample from moving between adjacent reaction chambers 20 when the sample reacts with reagents after being completely injected into the reaction chambers 20 provided in the body portion 100, and thus, reaction results of each reaction chambers 20 may be reliably secured without being mixed with reaction results of other reaction chambers 20.

[0058] The processing-solution test chamber portion 300 includes the test chamber 30 provided in the body portion 100 and accommodating a reaction reagent for testing a processing solution introduced through the processing-solution inlets 12 by allowing a reaction between the reaction agent and the processing solution. The reaction reagent is provided to detect whether a specific target genetic material is included in the processing solution. For example, the reaction reagent may be prepared in a form including a specific target genetic material to check whether the processing solution is contaminated with the specific target genetic material. The reaction reagent may be fixedly included in the test chamber 30.

[0059] In the present embodiment, the processing solution may be a solution such as a buffer for extracting nucleic acids from a sample. Before PCR is performed, a sample is pretreated to separate nucleic acids from a collected specimen. In the pretreatment, a predetermined buffer such as a lysis buffer is injected into a biological sample to break cell walls and release nucleic acids. The lysis buffer is a buffer for breaking cell walls. The sample in which nucleic acids are exposed is introduced into a reaction chamber. According to the present embodiment, the processing-solution test chamber portion is configured to check whether the processing solution used for pretreatment of the sample or the like is contaminated. Although the present embodiment has been described by way of example with respect to nucleic acid extraction using a lysis buffer, contamination of processing solutions used in other nucleic acid extraction methods, such as magnetic bead-based methods, column-type methods, and automated extraction methods, may also be checked.

[0060] According to an embodiment of the present disclosure, the processing solution may include not only a solution for pretreatment of a sample but also all processing solutions used for processing the sample. For example, the processing solution may include a sample dilution buffer such as DW or TE, a master mix, or the like. In this case, the master mix may be fixed to the reaction chambers 20 and the test chamber 30, or may be introduced into the reaction chambers 20 after an external mixing process.

[0061] As illustrated in FIG. 2, the test chamber 30 includes an inlet portion 31 through which a processing solution flows in, a well portion 32 that is recessed upward from the lower surface of the body portion 100 to allow a reaction between the processing solution and a reaction reagent, and an outlet portion 33 through which the processing solution flows out from the well portion 32. The reaction reagent that reacts with the processing solution may be fixedly accommodated in the well portion 32. According to the present embodiment, the test chamber 30 is formed substantially similar to the reaction chambers 20.

[0062] The processing-solution test chamber portion 300 may include at least one test chamber 30. When the processing-solution test chamber portion 300 includes a plurality of test chambers 30, the processing-solution test chamber portion 300 may include a flow path through which the processing solution is introduced, and the test chambers 30 may include a reaction reagent for detecting whether a specific target genetic material is included in the processing solution. In the embodiment shown in FIG. 2, four test chambers 30 are provided, and a processing-solution flow path 50 may connect the four test chambers 30 to each other. According to another embodiment as illustrated in FIG. 6, one test chamber 30 may be provided.

[0063] Referring to FIG. 2, a plurality of test chambers 30 are provided in the body portion 100. The test chambers 30 are connected to each other through the processing-solution flow path 50 such that the processing solution may flow in one direction. That is, the processing-solution flow path 50 and the test chambers 30 are connected in series to each other. The processing-solution inlets 12 through which the processing solution may be introduced are connected to both ends of the test chambers 30 that are connected in series to each other. A user may introduce a processing solution to be tested using any one of the processing-solution inlets 12.

[0064] According to the present embodiment, when a plurality of test chambers 30 are provided, the processing-solution flow path 50 for connecting the test chambers 30 to each other includes at least one curved portion in a section connecting one test chamber 30 and an adjacent test chamber 30. The processing-solution flow path 50 is preferably formed to have a size of less than 0.55 mm to prevent excessive pressure during movement of a processing solution and thus ensure smooth movement thereof. Each of the test chambers 30 is provided in the form of a groove extending in one direction.

[0065] The inlet portion 31 of each of the test chambers 30 includes an expanded portion 311 that gradually widens in a direction toward the test chamber 30. The expanded portion 311 is tapered such that when the processing solution flows into the test chamber 30, the processing solution is decelerated by a widening section and fills the internal space of the test chamber 30. The outlet portion 33 of each of the test chambers 30 includes a reduced portion 331 that gradually narrows in a direction away from the test chamber 30 for a processing solution flowing out from the test chamber 30. Due to the reduced portion 331, the flow speed of a processing solution increases, allowing the processing solution to easily move to an adjacent test chamber 30.

[0066] A bottom surface of the test chamber 30 is deeper than a bottom surface of the processing-solution flow path 50. The inlet portion 31 includes an inclined surface 34 connected to the bottom surface of the test chamber 30, and the outlet portion 33 similarly includes an inclined surface 34 connected to the bottom surface of the test chamber 30. The bottom surface of the test chamber 30 is deeper than the bottom surface of the processing-solution flow path 50, thereby forming sufficient space for fixing a reaction reagent, and because the reaction reagent is fixed to the deeper bottom surface, the reaction agent may have improved reactivity with a processing solution. When a fluorescent substance is used, the fluorescent substance may be more easily checked because the well portion 32 is deepened.

[0067] According to the present embodiment, the processing-solution flow path 50 is provided to connect a plurality of test chambers 30 to each other. When the test chambers 30 are referred to as N-th chambers (where N is a natural number) in an order adjacent to a processing-solution inlet 12, the processing-solution flow path 50 connects an N-th chamber and an (N+1)-th chamber. Here, the expression "in the order adjacent to a processing-solution inlet 12" refers to an order in which a processing solution passes through the test chambers 30 sequentially through the processing-solution flow path 50, starting from a reaction chamber 20 that the processing solution first reaches after being injected through the processing-solution inlet 12. According to the present embodiment, the processing-solution flow path 50 includes a third flow path 51, a fourth flow path 52, and a second connection flow path 53.

[0068] The third flow path 51 has one end connected to the N-th chamber and the other end extending toward the (N+1)-th chamber. The fourth flow path 52 has one end connected to the (N+1)-th chamber and the other end extending toward the N-th chamber. The second connection flow path 53 is provided between the third and fourth flow paths 51 and 52 to connect the third and fourth flow paths 51 and 52 to each other. That is, one end of the second connection flow path 53 is connected to the other end of the third flow path 51, and the other end of the second connection flow path 53 is connected to the end of the fourth flow path 52. According to the present embodiment, the third and fourth flow paths 51 and 52 include a bending portion 54 for changing the flow direction of a processing solution. At least one bending portion 54 may be provided.

[0069] According to the present embodiment, as illustrated in FIG. 3, the processing-solution flow path 50 is designed such that a processing solution changes direction three times while moving from the third flow path 51 to the second connection flow path 53, and three times while moving from the second connection flow path 53 to the fourth flow path 52. However, the number of bending portions 54 is not limited thereto and may vary. The bending portions 54 prevent the processing solution from moving between adjacent test chambers 30 when the processing solution reacts with reagents after being completely injected into the test chambers 30 provided in the body portion 100, and thus, reaction results of each test chamber 30 may be reliably secured without being mixed with reaction results of other test chambers 30. The test chambers 30 may include a plurality of reaction reagents for detecting specific target genetic materials.

[0070] According to the present embodiment, a cover portion 60 is coupled to the lower surface of the body portion 100.

[0071] The cover portion 60 is provided to close the sample test chamber portion 200 and the processing-solution test chamber portion 300, which are provided in the form of grooves recessed upward from the lower surface of the body portion 100. When the cover portion 60 is attached to the bottom surface of the body portion 100 and the body portion 100 is positioned such that the cover portion 60 faces downward, the cover portion 60 forms bottom surfaces of the sample test chamber portion 200 and the processing-solution test chamber portion 300. That is, when a sample and a processing solution injected through the sample inlets 11 and the processing-solution inlets 12 are delivered to the sample test chamber portion 200 and the processing-solution test chamber portion 300, bottom surfaces of the sample flow path 40 and the processing-solution flow path 50 are formed by the cover portion 60.

[0072] According to the present embodiment, the cover portion 60 is provided in the form of a film disposed on a lower side of the body portion 100 and attached to the bottom surface of the body portion 100, and heat for performing PCR may be transferred through the cover portion 60 from a heating device. The cover portion 60 may include a thin film, a metal having high heat transfer efficiency, or a polymer material containing nano-carbon to easily receive heat from the heating device. In addition, a sample and a processing solution may automatically move in the sample flow path 40 and the processing-solution flow path 50 without using separate power. An adhesive region R is provided on the bottom surface of the body portion 100 such that the cover portion 60 provided in the form of a film may be attached to the adhesive region R. The adhesive region R is wider than the sample flow path 40 and the processing-solution flow path 50, thereby preventing leakage when a processing solution or a sample is injected. The adhesive region R is formed at edges of the reaction chambers 20, the test chambers 30, the sample flow path 40, and the processing-solution flow path 50, and at an edge of the body portion 100. The adhesive region R may have a width of 0.8 mm to 1.0 mm. When the width of the adhesive region R is less than 0.8 mm, adhesion of the cover portion 60 decreases, and when the width exceeds 1.0 mm, a space for arrangement of the reaction chambers 20 is limited.

[0073] In addition, multi-functional PCR test chips 1001, 1002, and 1003 according to embodiments of the present disclosure are inserted into PCR equipment to perform PCR. The PCR equipment may include: a main body in which the chips 1001, 1002, and 1003 of the present disclosure are mounted; a heating unit provided inside the main body to provide heat to the chips 1001, 1002, and 1003; a light source for projecting light to the chips 1001, 1002, and 1003; a camera for capturing images in response to fluorescence generated when a reaction reagent and a sample react with each other; and an analysis unit for analyzing the images captured using the camera. The present disclosure relates to the chips 1001, 1002, and 1003, and thus a detailed description of the PCR equipment is omitted.(1) First embodiment

[0074] According to an embodiment of the present disclosure, a plurality of reaction chambers 20 and a plurality of test chambers 30 may be provided. For example, the plurality of reaction chambers 20 may be arranged adjacent to one another, and at least one test chamber 30 may be provided on each of both outermost sides of the plurality of reaction chambers 20. For example, in the multi-functional PCR test chip 1001 illustrated in FIG. 4, the sample test chamber portion 200 may include six reaction chambers 20, and the processing-solution test chamber portion 300 may include four test chambers 30. According to the present embodiment, one of the test chambers 30 is adjacent to one side of the six reaction chambers 20 arranged side by side, and the remaining three test chambers 30 are arranged side by side adjacent to an endmost one of the reaction chambers 20. In this case, the three test chambers 30 may be arranged adjacent to one another.

[0075] Specifically, the chip according to the embodiment shown in FIG. 4 may be utilized as follows. Reaction reagents capable of detecting six target genes (hereinafter referred to as target genes 1 to 6) are inserted together into a first test chamber 30, and reaction reagents each capable of detecting one target gene are inserted in second to seven reaction chambers 20, respectively.

[0076] That is, the reaction reagent capable of detecting the target gene 1 may be fixed to a second reaction chamber 20, the reaction reagent capable of detecting the target gene 2 may be fixed to a third reaction chamber 20, the reaction reagent capable of detecting the target gene 3 may be fixed to a fourth second reaction chamber 20, the reaction reagent capable of detecting the target gene 4 may be fixed to a fifth second reaction chamber 20, the reaction reagent capable of detecting the target gene 5 may be fixed to a sixth second reaction chamber 20, and the reaction reagent capable of detecting the target gene 6 may be fixed to a seventh second reaction chamber 20. In addition, the target genes 1 and 2 and the reaction reagents capable of detecting the target genes 1 and 2 may be fixed to an eighth test chamber 30, the target genes 3 and 4 and the reaction reagents capable of detecting the target genes 3 and 4 may be fixed to a ninth test chamber 30, and the target genes 5 and 6 and the reaction reagents capable of detecting the target genes 5 and 6 may be fixed to a tenth test chamber 30.

[0077] The positions of reaction reagents for detecting target genes may be varied depending on conditions, and the numbers of reaction chambers 20 and test chambers 30 may also be varied depending on designs. In this state, a processing solution (lysis buffer) is injected into the first test chamber 30 until the eighth to tenth test chambers 30 are filled with the processing solution, and a sample is injected into the second reaction chamber 20 until the seventh reaction chamber 20 is filled with the sample. Then, PCR is performed.

[0078] No reaction occurs in the first test chamber 30 because the processing solution is not contaminated with the target genes 1 to 6, verifying that the processing solution is not contaminated (NC). In addition, when amplification occurs in the chip inserted into the PCR equipment, successful amplification in the eighth to tenth test chambers 30 may verify that the PCR equipment and / or the reaction reagents function properly.

[0079] In addition, an internal control (IC) may be introduced into the reaction chambers 20 in order to verify whether nucleic acid extraction has been properly performed. The IC is used to verify an inherent gene that must be included in a specimen, and validity of nucleic acid extraction from a sample may be determined by determining, using the IC, whether extracted nucleic acids satisfy a quantitative criterion. The IC may be fixed to any one of the reaction chambers 20, or a user may directly introduce the IC into the reaction chambers 20 before PCR is performed.

[0080] In addition, two types of reaction reagents may be fixed to each reaction chamber to detect two target genetic materials in each reaction chamber. In this case, a total of twelve target genes may be detected in the embodiment shown in FIG. 4 because six reaction chambers 20 are provided. Correspondingly, reaction agents that reacts with 1 to 12 target genetic materials may be fixed to the first test chamber 30 to use the first test chamber 30 as a negative control (NC) for detecting whether the processing solution is contaminated with the twelve target genes. In addition, the eighth to tenth test chambers 30 may be used as a positive control (PC) for verifying the PCR equipment and the reagents.

[0081] As described above, PCR equipment, reaction reagents, and / or PCR processes may be verified using the test chambers 30 and the reaction chambers 20 In the state described above, when a processing solution or DW is introduced into the test chambers 30, no reaction with target genes accommodated in the test chambers 30 may verify that the processing solution is not contaminated (negative control: NC), smooth PCR amplification may verify that reaction agents function properly (positive control: PC).

[0082] In addition, a sample may be introduced into the reaction chambers 20 to test whether target genes are detected. That is, contamination of a processing solution, proper functioning of reagents, and testing of a sample may be performed using a single chip by injecting the processing solution into the test chambers 30 and the sample into the reaction chambers 20.(2) Second embodiment

[0083] FIG. 6 illustrates a multi-functional PCR test chip 1002 according to another embodiment of the present disclosure. As illustrated in FIG. 6, the present embodiment differs from the embodiment shown in FIG. 4 in the positions of sample inlets 11 and processing-solution inlets 12 on an upper side of a body portion 100. As illustrated in FIG. 7, the multi-functional PCR test chip 1002 of the present embodiment includes a plurality of reaction chambers 20 and one test chamber 30. Specifically, as illustrated in FIG. 8, a sample test chamber portion 200 and a processing-solution test chamber portion 300 are provided in a bottom surface of the body portion 100. The sample test chamber portion 200 includes nine reaction chambers 20, and the processing-solution test chamber portion 300 includes one test chamber 30.

[0084] Two types of reaction reagents for detecting two target genetic materials may be fixed to each of the reaction chambers 20. That is, in this case, a total of eighteen target genes may be detected. Correspondingly, reaction reagents capable of reacting with the eighteen target genes may be fixed to the test chamber 30 to detect whether a processing solution is contaminated with the eighteen target genes. Alternatively, a reaction reagent capable of detecting one target genetic material may be fixed to each of the reaction chambers 20.

[0085] In such a state, when no reaction is detected between the eighteen target genes and a processing solution or DW injected into the test chamber 30, the processing solution may be determined not to be contaminated, thereby enabling negative control (NC) testing. In addition, a sample may be injected into the reaction chambers 20 to test whether the target genes are detected. That is, when a test is performed by injecting a sample into the reaction chambers 20, contamination of a processing solution may be checked in the same chip.

[0086] In addition, an internal control (IC) may be injected into any one of the reaction chambers 20 verify whether nucleic acid extraction has been properly performed. The IC is a primer or probe capable of detecting an inherent gene possessed by a living organism, and may be used to check whether nucleic acid extraction from a specimen (sample) has been properly performed. Using the IC, the validity of nucleic acid extraction from the sample may be determined by assessing whether the extracted nucleic acids satisfy a quantitative criterion. The IC may be fixed to any one of the reaction chambers 20, or a user may directly inject the IC into the reaction chambers 20 before PCR is performed.

[0087] In the present embodiment, when a processing-solution test chamber portion 200 is configured by connecting a first chamber and a tenth chamber to each other, and a sample test chamber portion 200 is configured by connecting section to nineth chambers to each other, the tenth chamber may be used to function as a positive control (PC) as in the first embodiment.(3) Third embodiment

[0088] According to another embodiment of the present disclosure, a multi-functional PCR test chip 1003 may include a plurality of sample test chamber portions 200 and a plurality of processing-solution test chamber portions 300. When PCR is performed, any one of a plurality of sample test chamber portions 201 and 202 and any one of a plurality of processing-solution test chamber portions 301 and 302 may be used together as a set. That is, according to the present embodiment, a plurality of sets each including a combination of the sample test chamber portions 201 and 202 and the processing-solution test chamber portions 301 and 302 may be provided.

[0089] Referring to FIG. 9, the sample test chamber portions 200 include a first sample test chamber portion 201 including a plurality of reaction chambers 20 provided adjacent to one another, and a second sample test chamber portion 202 separated from the first sample test chamber portion 201 and including another plurality of reaction chambers 20 provided adjacent to one another. In addition, the processing-solution test chamber portion 300 may include at least one first test chamber 301 adjacent to the first sample test chamber portion 201, and at least one second test chamber 302 adjacent to the second sample test chamber portion 202. According to the present embodiment, the first sample test chamber portion 201 and the second sample test chamber portion 202 are disposed adjacent to each other between the first test chamber 301 and the second test chamber 302.

[0090] According to the present embodiment, as illustrated in FIG. 9, the first test chamber 301 and the first sample test chamber portion 201 form one set, and the second sample test chamber portion 202 and the second test chamber 302 form another set.

[0091] Specifically, the first sample test chamber portion 201 includes four reaction chambers 20 arranged adjacent to one another and connected to each other through a sample flow path 40. The second sample test chamber portion 202 is not connected to the first sample test chamber portion 202, and includes four reaction chambers 20 arranged adjacent to one another. The second sample test chamber portion 202 also includes a sample flow path 40 as a connection path.

[0092] The number of reaction chambers 20 of each of the first and second sample test chamber portions 201 and 202 is not limited to the four described above and may vary. Referring to FIG. 9, the first test chamber 301 is disposed adjacent to a left outermost side of the first sample test chamber portion 201, and the second test chamber 302 is disposed adjacent to a right outermost side of the second sample test chamber portion 202. The first test chamber 301 and the first sample test chamber portion 201 are formed symmetrically with the second test chamber 302 and the second sample test chamber portion 202. Alternatively, the first test chamber 301 may be provided on a right outermost side of the first sample test chamber portion 201, or the second test chamber 302 may be provided on a left outermost side of the second sample test chamber portion 202.

[0093] Two processing-solution inlets 12 are provided in each of the first and second test chambers 301 and 302. Two sample inlets 11 are provided at both ends of each of the first sample test chamber portion 201 and the second sample test chamber portion 202. According to the present embodiment, the first test chamber 301 may be used to test contamination of a processing solution or DW used to process a first sample, and the first sample test chamber portion 201 may be used to test whether a target gene is detected in the first sample introduced into the reaction chambers 20. In addition, the second test chamber 302 may be used to test contamination of a processing solution or DW used to process a second sample, and the second sample test chamber portion 202 may be used to test whether a target gene is detected in the second sample introduced into the reaction chambers 20.

[0094] Referring again to FIG. 9, two types of reaction reagents capable of detecting two target genetic materials may be fixed to each of second to fifth reaction chambers 20 of the first sample test chamber portion 201. That is, in this case, a total of eight target genes may be detected. Correspondingly, reaction reagents capable of reacting with the eight target genetic materials may be fixed to the first test chamber 301 to detect whether the processing solution for processing the first sample is contaminated with the eight target genes. Alternatively, a reaction reagent for detecting one target genetic material may be fixed to each of the reaction chambers 20 of the first sample test chamber portion 201.

[0095] In such a state, when the processing solution or DW for processing the first sample is introduced into the first test chamber 301, and no reaction is detected between the processing solution or DW and the eight target genes, the processing solution is determined not to be contaminated such that the first test chamber may function as a negative control (NC). In addition, the first sample may be introduced into the first sample test chamber portion 201 to test whether target genes are detected.

[0096] That is, when a test is performed by introducing the first sample into the reaction chambers 20, contamination of the processing solution for processing the first sample may be checked in the same chip. In addition, an internal control (IC) may be introduced into any one of the reaction chambers 20 to verify whether nucleic acid extraction has been properly performed. As described above, using the IC, validity of nucleic acid extraction from a sample may be determined by assessing whether the extracted nucleic acids satisfy a quantitative criterion. The IC may be fixed to any one of the reaction chambers 20 of the first sample test chamber portion 201, or a user may directly introduce the IC into the reaction chambers 20 before PCR is performed.

[0097] Alternatively, in the present embodiment, first and fifth chambers may be connected to each other to form the first test chamber 301, and second to fourth chambers may be connected to each other to form the first sample test chamber portion 201. In this case, the fifth chamber may be used to function as a positive control (PC) as in the first embodiment.

[0098] Similarly to the first sample test chamber portion 201, in the second sample test chamber portion 202, two kinds of reaction reagents for detecting two target genetic materials may be fixed to each of sixth to nineth reaction chambers 20. That is, in this case, a total of eight target genes may be detected. The eight target genes may be the same as the eight target genes of the first sample test chamber portion 201. reaction reagents capable of reacting with the eight target genetic materials may be fixed to the second test chamber 302 to detect whether the processing solution for processing the second sample is contaminated with the eight target genes, Alternatively, a reaction reagent for detecting one target genetic material may be fixed to each of the reaction chambers 20 of the second sample test chamber portion 202.

[0099] In such a state, when the processing solution or DW for processing the second sample is introduced into the second test chamber 302, and no reaction is detected between the processing solution or DW and the eight target genes, the processing solution is determined not to be contaminated such that the first test chamber may function as a negative control (NC). In addition, the second sample may be introduced into the second sample test chamber portion 202 to test whether target genes are detected. That is, when a test is performed by introducing the second sample into the reaction chambers 20, contamination of the processing solution for processing the second sample may be checked in the same chip.

[0100] In addition, an internal control (IC) may be introduced into any one of the reaction chambers 20 to verify whether nucleic acid extraction has been properly performed. As described above, using the IC, validity of nucleic acid extraction from a sample may be determined by assessing whether the extracted nucleic acids satisfy a quantitative criterion. The IC may be fixed to any one of the reaction chambers 20 of the second sample test chamber portion 202, or a user may directly introduce the IC into the reaction chambers 20 before PCR is performed.

[0101] Alternatively, in the present embodiment, sixth and tenth chambers may be connected to each other to form the second test chamber 302, and seventh to nineth chambers may be connected to each other to form the second sample test chamber portion 202. In this case, the sixth chamber may be used to function as a positive control (PC) as in the first embodiment.

[0102] As described above, the present embodiment provides a chip capable of simultaneously testing different samples. According to the present embodiment, two samples may be tested with one chip, and in this case, contamination of processing solutions for processing the two samples may be simultaneously checked. The numbers of reaction chambers and test chambers or the types and numbers of reaction reagents fixed to the reaction chambers and the test chambers may be varied depending on target genetic materials to be detected.

[0103] Through the processes described above, the present disclosure enables multiplex testing by detecting multiple types of target genes with a single injection of a sample, and also enables contamination of a processing solution, such as a buffer used to extract nucleic acids from the sample, to be checked together on a single chip. That is, a sample and a processing solution may be simultaneously tested owing to a configuration in which a sample test chamber portion, including a plurality of reaction chambers into which the sample is introduced, is separated from a processing-solution test chamber portion that functions as a negative control (NC) by receiving the processing solution or the like to check whether the processing solution is contaminated.

[0104] In addition, because the processing-solution test chamber portion includes a plurality of test chambers for checking whether reaction reagents function properly (positive control (PC)), testing of a sample, testing of contamination of a processing solution, and checking whether reaction reagents function properly may be simultaneously performed.

[0105] In addition, the present disclosure aims to enable simultaneous multiplex diagnosis with high sensitivity and specificity. In multiplex PCR diagnosis of the related art, a plurality of primer probes for detecting multiple targets are disposed in the same space, leading to interference, competitive reactions, and variations in reaction characteristics, thereby degrading sensitivity and specificity. To address this problem, target detection regions are spatially separated from each other to eliminate mutual interference and competitive reactions. In addition, because reaction reagents react with a sample in a state in which the reaction agents are fixed to reaction chambers, sensitivity may be improved when checking test results.

[0106] While preferred embodiments of the present disclosure have been described above in detail, the present disclosure is not limited to the embodiments, and various modifications may be made without departing from the scope of the present disclosure.

Examples

first embodiment

(1) First embodiment

[0074]According to an embodiment of the present disclosure, a plurality of reaction chambers 20 and a plurality of test chambers 30 may be provided. For example, the plurality of reaction chambers 20 may be arranged adjacent to one another, and at least one test chamber 30 may be provided on each of both outermost sides of the plurality of reaction chambers 20. For example, in the multi-functional PCR test chip 1001 illustrated in FIG. 4, the sample test chamber portion 200 may include six reaction chambers 20, and the processing-solution test chamber portion 300 may include four test chambers 30. According to the present embodiment, one of the test chambers 30 is adjacent to one side of the six reaction chambers 20 arranged side by side, and the remaining three test chambers 30 are arranged side by side adjacent to an endmost one of the reaction chambers 20. In this case, the three test chambers 30 may be arranged adjacent to one another.

[0075]Specifically, the ...

second embodiment

(2) Second embodiment

[0083]FIG. 6 illustrates a multi-functional PCR test chip 1002 according to another embodiment of the present disclosure. As illustrated in FIG. 6, the present embodiment differs from the embodiment shown in FIG. 4 in the positions of sample inlets 11 and processing-solution inlets 12 on an upper side of a body portion 100. As illustrated in FIG. 7, the multi-functional PCR test chip 1002 of the present embodiment includes a plurality of reaction chambers 20 and one test chamber 30. Specifically, as illustrated in FIG. 8, a sample test chamber portion 200 and a processing-solution test chamber portion 300 are provided in a bottom surface of the body portion 100. The sample test chamber portion 200 includes nine reaction chambers 20, and the processing-solution test chamber portion 300 includes one test chamber 30.

[0084]Two types of reaction reagents for detecting two target genetic materials may be fixed to each of the reaction chambers 20. That is, in this case...

third embodiment

(3) Third embodiment

[0088]According to another embodiment of the present disclosure, a multi-functional PCR test chip 1003 may include a plurality of sample test chamber portions 200 and a plurality of processing-solution test chamber portions 300. When PCR is performed, any one of a plurality of sample test chamber portions 201 and 202 and any one of a plurality of processing-solution test chamber portions 301 and 302 may be used together as a set. That is, according to the present embodiment, a plurality of sets each including a combination of the sample test chamber portions 201 and 202 and the processing-solution test chamber portions 301 and 302 may be provided.

[0089]Referring to FIG. 9, the sample test chamber portions 200 include a first sample test chamber portion 201 including a plurality of reaction chambers 20 provided adjacent to one another, and a second sample test chamber portion 202 separated from the first sample test chamber portion 201 and including another plural...

Technical Field

[0001] The present disclosure relates to a multi-functional PCR test chip for reacting a sample with a reaction reagent to perform a PCR test. The reaction reagent is fixed to the chip, the sample to be tested is introduced into the chip, and a PCR test is performed by checking whether the sample reacts with the reaction agent.Background Art

[0002] Polymerase chain reaction (hereinafter referred to as PCR) refers to a method for amplifying a specific target genetic material to be detected, that is, a technique for amplifying a small amount of genetic material to obtain a large amount of genetic material having the same base sequence as the original genetic material. PCR is used to amplify human nucleic acids to diagnose various types of genetic diseases or is applied to nucleic acids of bacteria, viruses, or fungi to diagnose infectious diseases.

[0003] In general, PCR is performed by repeatedly carrying out the following three steps. The three steps include: 1) a denaturing step of heating a sample solution containing double-stranded DNA to a specific temperature, for example, about 95 °C, to separate the double-stranded DNA into single-stranded DNA; 2) an annealing step of, after the denaturing step, providing the sample solution with an oligonucleotide primer having a sequence complementary to a specific base sequence to be amplified, and cooling the sample solution together with the single-stranded DNA to a specific temperature, for example, 55 °C, to bind the primer to the specific base sequence of the single-stranded DNA to form a partial DNA-primer complex; and 3) an extension (or amplification) step of, after the annealing step, maintaining the sample solution at an activation temperature of a DNA polymerase, for example, 72 °C, to form double-stranded DNA using the DNA polymerase based on the primer of the partial DNA-primer complex. A target nucleic acid having the specific base sequence may be exponentially amplified by repeating these three steps multiple times. However, in the case of RNA, a step of reverse-transcribing the RNA into DNA may be performed prior to the steps 1) to 3).

[0004] To perform such PCR, a PCR chip equipped with a chamber capable of accommodating a reagent and a sample and a PCR device configured to heat and cool the PCR chip to induce an amplification reaction and to measure a result thereof are required. As devices for amplifying target nucleic acids, various PCR devices have been developed.

[0005] In the related art, after completion of PCR, only qualitative analysis of amplified target nucleic acids through a separate electrophoresis process was possible. However, real-time PCR devices (that is, rPCR devices) have recently been developed, which enable quantitative analysis of target nucleic acids by detecting fluorescence intensity proportional to the concentration of amplified genetic material using an optical detection system. In addition, portable real-time PCR devices have been developed by miniaturizing large PCR devices used in laboratories, thereby enabling PCR at a sample collection site. Portable PCR devices are characterized by the use of PCR chips (or biochips or micro-chips) and cameras or digital cameras instead of large and complex optical detection systems.

[0006] The present disclosure relates to a PCR test chip that enhances sensitivity when detecting the intensity of fluorescence emitted during a reaction between a reagent and a sample, enables simultaneous detection of multiple target genes, and enables simultaneous checking for contamination of a processing solution such as a buffer used for extracting nucleic acids from a sample.Disclosure of Invention Technical Problem

[0007] In the related art, a processing solution (buffer solution) is used to extract nucleic acids from a sample, and nucleic acids included in the buffer solution are introduced into a chip to test whether a reaction with reaction reagents occurs. However, there is an inconvenience in that contamination of the processing solution is not separately checked, and an additional procedure is performed to verify such contamination.

[0008] The present disclosure is provided to address the above-described problems. An objective of the present disclosure is to provide a PCR test chip in which reaction reagents that react with a sample are fixed to reaction chambers, thereby enhancing sensitivity in checking test results and enabling contamination of a processing solution, such as a buffer used to extract nucleic acids from a sample, to be checked together.Solution to Problem

[0009] According to an embodiment of the present disclosure, a multi-functional PCR test chip includes: a body portion including a light-transmissive material, a sample inlet through which a sample is introduced, and a processing-solution inlet through which a processing solution for processing the sample is introduced; a sample test chamber portion provided in the body portion and including a reaction chamber, the reaction chamber accommodating a reaction reagent for detecting a target genetic material and in which the sample introduced through the sample inlet reacts with the reaction reagent; and a processing-solution test chamber portion provided in the body portion and including a test chamber, the test chamber accommodating the reaction reagent and in which the processing solution introduced through the processing-solution inlet reacts with the reaction reagent.

[0010] In addition, the reaction chamber may preferably include: an inlet portion through which the sample flows in; a reaction well portion in which the sample reacts with the reaction reagent, the reaction well portion being recessed upward from a lower surface of the body portion; and an outlet portion through which the sample flows out from the reaction well portion.

[0011] In addition, preferably, the inlet portion and the outlet portion of the reaction chamber may be inclined.

[0012] In addition, preferably, a plurality of reaction chambers may be provided in the body portion, and the plurality of reaction chambers may be connected to each other through a sample flow path to allow the sample to flow in one direction.

[0013] In addition, preferably, the inlet portion may include an expanded portion that widens in a direction in which the sample enters the reaction well portion, and the outlet portion may include a reduced portion that narrows in a direction in which the sample flows out from the reaction well portion.

[0014] In addition, preferably, when the plurality of reaction chambers are referred to as N-th chambers (where N is a natural number) in order adjacent to the sample inlet, the sample flow path may connect an N-th chamber and an (N+1)-th chamber to each other, wherein the sample flow path may include: a first flow path having one end connected to the N-th chamber; a second flow path having another end connected to the (N+1)-th chamber; and a first connection flow path provided between the first flow path and the second flow path and connecting the first flow path and the second flow path to each other, wherein the first flow path and the second flow path may include at least one bending portion to change a flow direction of the sample.

[0015] In addition, preferably, a cover portion may be coupled to a lower surface of the body portion to close the sample test chamber portion and the processing-solution test chamber portion that are provided as grooves recessed upward from the lower surface of the body portion, and in a state in which the body portion is positioned with the cover portion facing downward, the cover portion may form bottom surfaces of the sample test chamber portion and the processing-solution test chamber portion.

[0016] In addition, preferably, the cover portion may be provided as a film disposed on a lower side of the body portion and attached to the lower surface of the body portion to transfer heat from a heating device for performing PCR.

[0017] In addition, preferably, the test chamber may include a plurality of test chambers.

[0018] In addition, preferably, the plurality of test chambers may be connected to each other through a processing-solution flow path to allow the processing solution to flow in one direction.

[0019] In addition, preferably, when the plurality of test chambers are referred to as M-th chambers (where M is a natural number) in order adjacent to the processing-solution inlet, the processing-solution flow path may connect an M-th chamber and an (M+1)-th chamber to each other, wherein the processing-solution flow path may include: a third flow path having one end connected to the M-th chamber; a fourth flow path having another end connected to the (M+1)-th chamber; and a second connection flow path provided between the third flow path and the fourth flow path and connecting the third flow path and the fourth flow path to each other, wherein the third flow path and the fourth flow path may include at least one bending portion to change a flow direction of the sample.

[0020] In addition, preferably, the plurality of reaction chambers may be arranged adjacent to one another, and the test chamber may include a plurality of test chambers, at least one of the plurality of test chambers being provided on each of both outermost sides of the plurality of reaction chambers arranged adjacent to one another.

[0021] In addition, preferably, the sample test chamber portion may include: a first sample test chamber portion including a plurality of reaction chambers arranged adjacent to each other; and a second sample test chamber portion provided apart from the first sample test chamber portion and including another plurality of reaction chambers arranged adjacent to each other, wherein the processing-solution test chamber portion may include at least one first test chamber adjacent to the first sample test chamber portion and at least one second test chamber adjacent to the second sample test chamber portion.

[0022] In addition, preferably, the first sample test chamber portion and the second sample test chamber portion may be arranged adjacent to each other between the first test chamber and the second test chamber.

[0023] In addition, preferably, reaction reagents for detecting at least two different target genes may be fixed to the reaction chamber and the test chamber to enable multiplex diagnosis for samples.Advantageous Effects of Invention

[0024] An embodiment of the present disclosure provides a PCR test chip in which reaction reagents that react with a sample are fixed to reaction chambers, thereby improving sensitivity in checking test results and enabling contamination of a processing solution, such as a buffer used to extract nucleic acids from a sample, to be checked together.

[0025] Specifically, the present disclosure aims to enable simultaneous multiplex diagnosis with high sensitivity and specificity. In multiplex PCR diagnosis of the related art, a plurality of primer probes for detecting multiple targets are disposed in the same space, leading to interference, competitive reactions, and variations in reaction characteristics, thereby degrading sensitivity and specificity. To address this problem, target detection regions are spatially separated from each other to eliminate mutual interference and competitive reactions.

[0026] In addition, unlike multiplex PCR diagnosis of the related art, there is no need for multiple preparations of reaction solutions, and because the reaction solutions are connected to each other through a single flow path, a reaction may be performed simply and rapidly with a single preparation and injection of reaction solutions.

[0027] Further, although the reaction solutions are connected to each other through a single flow path, diffusion or interference between target spaces are prevented by microfluidic techniques, thereby improving the accuracy of test results.

[0028] Further, a flow path for contamination checking (NC: negative control) and a flow path for verification of equipment and reagents (PC: positive control) are separately designed, thereby providing an effect of enabling not only inspection of a sample but also NC testing and / or PC testing to be performed together.

[0029] Further, a sample flow path, through which reaction chambers are connected to each other, and a processing-solution flow path, through which test chambers are connected to each other, are provided with bending portions, thereby effectively preventing a processing solution or a sample from moving from one reaction chamber or test chamber to an adjacent reaction chamber or test chamber when the processing solution or sample reacts with reaction reagents. As a result, the accuracy of test results may be improved.

[0030] Further, because the reaction chambers or test chambers include recessed wells for reaction reagents, reaction may occur in the wells, thereby improving visibility. In addition, each of the reaction chambers or test chambers is provided with an expanded portion at an inlet and a reduced portion at an outlet, and thus, a sample or processing solution may be rapidly dispensed (diffused).Brief Description of Drawings

[0031] FIG. 1 is a perspective view illustrating a multi-functional PCR test chip according to an embodiment of the present disclosure, FIG. 2 is a bottom perspective view of FIG. 1, FIG. 3 is an enlarged view of FIG. 2, FIG. 4 is a view illustrating functions of a reaction chamber and a test chamber of FIG. 1, FIG. 5 is a cross-sectional view of FIG. 1, FIG. 6 is a perspective view illustrating a multi-functional PCR test chip according to another embodiment of the present disclosure, FIG. 7 is a bottom perspective view of FIG. 6, FIG. 8 is a view illustrating functions of a reaction chamber and a test chamber of FIG. 7, and FIG. 9 is a view illustrating a multi-functional PCR test chip according to still another embodiment of the present disclosure. Mode for the Invention

[0032] Hereinafter, various embodiments of the present disclosure will be described with reference to the accompanying drawings. Various embodiments of the present disclosure may be subject to various modifications and may have various forms, and specific embodiments are illustrated in the drawings and described in detail. However, this is not intended to limit the various embodiments of the present disclosure to specific forms, and all modifications and / or equivalents or substitutes within the spirit and technical scope of the various embodiments should be considered as being included in the scope of the present disclosure. In the drawings, like reference numerals denote like elements.

[0033] In various embodiments of the present disclosure, expressions such as "include" or "may include" specify the presence of the disclosed corresponding functions, operations, or elements, and do not preclude the presence of one or more additional functions, operations, or elements. Furthermore, in various embodiments of the present disclosure, terms such as "include" or "have" are intended to specify the presence of features, numbers, steps, operations, elements, components, or combinations thereof described in the specification, and do not to preclude the presence or addition of one or more other features, numbers, steps, operations, elements, components, or combinations thereof.

[0034] It should be understood that when an element is referred to as being "connected to" another element, the element may be directly connected to the other element, or any other element may be interposed between the two elements. In contrast, it should be understood that when an element is referred to as being "directly connected to" or "directly coupled to" another element, there is no other element between the two elements.

[0035] Terms used in various embodiments of the present disclosure are only for describing specific embodiments and are not intended to limit the various embodiments of the present disclosure. The terms of a singular form may include plural forms unless referred to the contrary.

[0036] Unless defined otherwise, all terms used herein, including technical and scientific terms, have the same meaning as those commonly understood by a person skilled in the art to which various embodiments of the present disclosure pertain.

[0037] Terms defined in generally used dictionaries should be interpreted as having meanings consistent with the context of the related art, and unless explicitly defined in various embodiments of the present disclosure, should not be interpreted as having idealized or excessively formal meanings.

[0038] The present disclosure relates to a chip used in a PCR device. The chip accommodates a reaction reagent, and after a sample is injected into the chip, a target gene is detected using the PCR device. The present disclosure provides a structure in which a portion having a plurality of reaction chambers into which a sample is injected is separated from a chamber portion for testing contamination of an introduced processing solution, a reaction reagent, or the like to enable negative control (NC) testing and / or positive control (PC) testing, thereby enabling simultaneous inspection of the sample and the processing solution.

[0039] In the present specification, the term "polymerase chain reaction (PCR)" refers to a reaction for amplifying a specific target nucleic acid molecule by using a thermostable DNA polymerase. For PCR, a reaction mixture including, in addition to a DNA polymerase, primers (forward primers and reverse primers), which are oligonucleotides capable of hybridizing specifically with a target nucleic acid, a deoxynucleotide triphosphate (dNTP) mixture, and divalent ions such as Mg 2+< may be used.

[0040] A "primer" is used to initiate a PCR process and refers to an oligonucleotide or polynucleotide that hybridizes complementarily with a template DNA. As primers for a PCR process, a forward primer (or sense primer) selected from a sense strand having the same direction as the genetic code progression of a nucleic acid molecule to be amplified, and a reverse primer (or antisense primer) selected from an antisense strand complementary to the sense strand, may be used as a pair.

[0041] The term "sample" refers to a genetic material to be amplified or a biological solution containing such genetic material. The term "reaction reagent" refers to a reagent that is used to detect a target genetic material and may include primers or the like. Primers may include a pair of primers having a length of 15 bp to 30 bp and capable of binding to both ends of a specific region of a target gene. In addition, an enzyme that does not lose activity even at a high temperature of 90 °C or higher is used as a DNA polymerase.

[0042] Hereinafter, preferred embodiments of the present disclosure will be described in detail with reference to the accompanying drawings. According to an embodiment of the present disclosure, a multi-functional PCR test chip 1001 includes a body portion 100, a sample test chamber portion 200, and a processing-solution test chamber portion 300.

[0043] The body portion 100 may include a light-transmissive material to detect a fluorescent substance by using an optical detection device. As illustrated in FIG. 1, according to the present embodiment, the body portion 100 may include a transparent polymer material. The body portion 100 may have a rectangular shape. The body portion 100 may have various shapes such as a circular shape or a square shape. According to the present embodiment, protruding walls 13 are formed on opposite long sides of the rectangular shape of the body portion 100 such that a user may easily grip the body portion 100. Reinforcing protrusions 14 extending in a vertical direction are formed on inner sides of the protruding walls 13. The reinforcing protrusions 14 prevent an injected material from bending when the body portion 100 is formed by injection molding, and improve the rigidity of the body portion 100. In addition, during product manufacture or in the field, the reinforcing protrusions 14 allow chips to be stacked in a vertical direction, and in this case, the reinforcing protrusions 14 may function as supports for supporting an upper chip. In addition, grooves are formed in outer peripheral surfaces of the protruding walls 13 such that a user may easily grip the body portion 100.

[0044] In addition, as illustrated in FIGS. 1 and 2, the body portion 100 includes sample inlets 11 into which a sample is introduced, and processing-solution inlets 12 into which a processing solution for processing the sample is introduced. Flow paths 40 and 50 and a plurality of chambers 20 and 30, all of which are shaped like grooves, are formed in the body portion 100. A sealing film is attached to the body portion 100, and together with the flow paths 40 and 50 and the chambers 20 and 30, the sealing film forms flow paths through which a sample and a processing solution move.

[0045] The sample test chamber portion 200, which is provided in the body portion 100, includes the reaction chamber 20 in which a reaction reagent for detecting a target genetic material is accommodated such that a sample introduced through the sample inlets 11 may react with the reaction reagent. According to the present embodiment, the sample test chamber portion 200 may include a flow path extending in one direction and a plurality of reaction chambers 20 provided along the flow path.

[0046] As illustrated in FIG. 2, each of the reaction chambers 20 includes an inlet portion 21 through which a sample flows in, a reaction well portion 22 recessed upward from a lower surface of the body portion 100 to allow a reaction between the sample and a reaction reagent, and an outlet portion 23 through which the sample flows out from the reaction well portion 22. Each of the reaction chambers 20 may include a reaction reagent that reacts with a sample. Specifically, the reaction reagent that reacts with the sample may be fixed to the reaction well portion 22.

[0047] According to the present embodiment, a plurality of reaction chambers 20 are provided in the body portion 100. The reaction chambers 20 are connected to each other through a sample flow path 40 to allow a sample to flow in one direction. That is, the sample flow path 40 and the reaction chambers 20 are connected in series to each other. The sample inlets 11, which are provided to receive a sample, are connected to both ends of the reaction chambers 20 that are connected in series to each other. When testing a sample, a user may inject the sample through any one of the sample inlets 11.

[0048] The reaction reagent may include a primer or the like. In addition, the reaction reagent may further include a fluorescent dye. Depending on a PCR method, the fluorescent dye may be injected into the chip together with a sample. The fluorescent dye emits light in a specific wavelength band during a DNA extension (or amplification) step. At least two types of reaction reagents may be provided to detect different target genes, and different reaction reagents may be accommodated in the reaction chambers 20. Because a plurality of reaction chambers 20 are provided, different reaction reagents may be fixed to the reaction chambers 20 to enable detection of different target genes with a single injection of a sample.

[0049] For example, in an embodiment, six reaction chambers 20 may be provided. When reaction reagents that each detect two types of specific target genes are fixed to the reaction chambers 20, respectively, a total of twelve target genes may be detected. In this case, the reaction reagents may be fixed to the reaction chambers 20 in a state in which each of the reaction reagents includes a fluorescent dye. Alternatively, the fluorescent dye may be injected into the chip together with a sample. Fluorescent dyes may be provided by fixing a fluorescent die together with different reaction reagents, injecting a first die together with a sample, or injecting different first and second dies together with a sample. The fluorescent dyes respond to light of a specific wavelength, and the first and second dyes may generate fluorescence with respect to light of different wavelengths. When amplification is performed by a reaction with a target gene in each the reaction chambers 20, a reaction in the reaction chamber 20 may be identified by detecting emission of light. When the first dye is injected identically into six chambers 20, six types of multiplex diagnosis may be possible at once by projecting light of a single wavelength to which the first dye responds. When the first dye and the second dye are simultaneously injected into each of the reaction chambers 20, twelve types of multiplex diagnosis may be possible at once by projecting multi-wavelength light having two different wavelengths.

[0050] The reaction reagents are fixed to the reaction chambers 20, and the chip in which the reaction reagents are fixed may be transported to a place at which PCR testing is required and used as needed. The reaction reagents may be fixed to the reaction chambers 20 by a method such as natural drying, vacuum drying, or freeze-drying. In addition, surfaces of the reaction reagents fixed to the reaction chambers 20 may be coated with a waterproof and adhesive coating agent.

[0051] The coating agent may be fixed together with reagents (primers, probes, or premixes) by using a biopolymer material that does not affect properties of the reagents or PCR results. The coating agent may maintain a solid state at room temperature and may melt at a temperature equal to or higher than a predetermined temperature. For example, the coating agent may melt at 45 °C or higher. When the chip into which a sample is injected is heated during a PCR process, the coating agent may melt, allowing a reaction between the sample and a reaction reagent. The coating agent may include any one of wax, agarose, paraffin, collagen, chitosan, and gelatin, or a mixture thereof. The coating agent may be fixed to a reaction chamber in a state in which the coating agent is mixed with a reaction reagent. Even in this case, the reaction reagent accommodated in the reaction chamber may maintain a solid state at room temperature, and when the chip into which the sample is injected is heated during a PCR process, the coating agent may melt to allow a reaction between the sample and the reaction reagent.

[0052] According to the present embodiment, the sample flow path 40, through which a sample is transported, connects the reaction chambers 20 to each other and includes at least one curved portion in a section connecting adjacent reaction chambers 20. The sample flow path 40 preferably has a size of less than 0.55 mm to prevent excessive pressure during movement of a sample and thus ensure smooth movement thereof. Each of the reaction chambers 20 is provided in the form of a groove extending in one direction.

[0053] The inlet portion 21 of each of the reaction chambers 20 includes an expanded portion 211 that gradually widens in a direction toward the reaction chamber 20. The expanded portion 211 is tapered such that when a sample flows into the reaction chamber 20, the sample is decelerated by a widening section and fills the internal space of the reaction chamber 20. The outlet portion 23 of each of the reaction chambers 20 includes a reduced portion 231 that gradually narrows in a direction away from the reaction chamber 20 for a sample flowing out from the reaction chamber 20. Owing to the reduced portion 231, the flow speed of a sample may increase, and thus, the sample may easily flow to an adjacent reaction chamber 20.

[0054] A bottom surface of the reaction chamber 20 is deeper than a bottom surface of the sample flow path 40. The inlet portion 21 includes an inclined surface 24 connected to the bottom surface of the reaction chamber 20, and the outlet portion 23 similarly includes an inclined surface 24 connected to the bottom surface of the reaction chamber 20. The bottom surface of the reaction chamber 20 is deeper than the bottom surface of the sample flow path 40, thereby forming a sufficient space for fixing a reaction reagent, and because the reaction reagent is fixed to the deeper bottom surface, the reaction agent may have improved reactivity with the sample. When a fluorescent substance is used, the fluorescent substance may be more easily checked because the reaction well portion 22 is deepened.

[0055] According to the present embodiment, the sample flow path 40 is provided to connect a plurality of reaction chambers 20 to each other. When the reaction chambers 20 are referred to as N-th chambers (where N is a natural number) in an order adjacent to a sample inlet 11, the sample flow path 40 connects an N-th chamber and an (N+1)-th chamber. Here, the expression "in the order adjacent to a sample inlet 11" refers to an order in which a sample passes through the reaction chambers 20 sequentially through the sample flow path 40, starting from a reaction chamber 20 that the sample first reaches after being injected through the sample inlet 11. According to the present embodiment, the sample flow path 40 includes a first flow path 41, a second flow path 42, and a first connection flow path 43.

[0056] The first flow path 41 has one end connected to the N-th chamber and the other end extending toward the (N+1)-th chamber. The second flow path 42 has one end connected to the (N+1)-th chamber and the other end extending toward the N-th chamber. The first connection flow path 43 is provided between the first and second flow paths 41 and 42 to connect the first and second flow paths 41 and 42 to each other. That is, one end of the first connection flow path 43 is connected to the other end of the first flow path 41, and the other end of the first connection flow path 43 is connected to the end of the second flow path 42. According to the present embodiment, the first and second flow paths 41 and 42 include a bending portion 44 for changing the flow direction of a sample. At least one bending portion 44 may be provided.

[0057] According to the present embodiment, as illustrated in FIG. 3, the sample flow path 40 is designed such that a sample changes direction three times while moving from the first flow path 41 to the first connection flow path 43, and three times while moving from the first connection flow path 43 to the second flow path 42. However, the number of bending portions 44 is not limited thereto and may vary. The bending portions 44 prevent a sample from moving between adjacent reaction chambers 20 when the sample reacts with reagents after being completely injected into the reaction chambers 20 provided in the body portion 100, and thus, reaction results of each reaction chambers 20 may be reliably secured without being mixed with reaction results of other reaction chambers 20.

[0058] The processing-solution test chamber portion 300 includes the test chamber 30 provided in the body portion 100 and accommodating a reaction reagent for testing a processing solution introduced through the processing-solution inlets 12 by allowing a reaction between the reaction agent and the processing solution. The reaction reagent is provided to detect whether a specific target genetic material is included in the processing solution. For example, the reaction reagent may be prepared in a form including a specific target genetic material to check whether the processing solution is contaminated with the specific target genetic material. The reaction reagent may be fixedly included in the test chamber 30.

[0059] In the present embodiment, the processing solution may be a solution such as a buffer for extracting nucleic acids from a sample. Before PCR is performed, a sample is pretreated to separate nucleic acids from a collected specimen. In the pretreatment, a predetermined buffer such as a lysis buffer is injected into a biological sample to break cell walls and release nucleic acids. The lysis buffer is a buffer for breaking cell walls. The sample in which nucleic acids are exposed is introduced into a reaction chamber. According to the present embodiment, the processing-solution test chamber portion is configured to check whether the processing solution used for pretreatment of the sample or the like is contaminated. Although the present embodiment has been described by way of example with respect to nucleic acid extraction using a lysis buffer, contamination of processing solutions used in other nucleic acid extraction methods, such as magnetic bead-based methods, column-type methods, and automated extraction methods, may also be checked.

[0060] According to an embodiment of the present disclosure, the processing solution may include not only a solution for pretreatment of a sample but also all processing solutions used for processing the sample. For example, the processing solution may include a sample dilution buffer such as DW or TE, a master mix, or the like. In this case, the master mix may be fixed to the reaction chambers 20 and the test chamber 30, or may be introduced into the reaction chambers 20 after an external mixing process.

[0061] As illustrated in FIG. 2, the test chamber 30 includes an inlet portion 31 through which a processing solution flows in, a well portion 32 that is recessed upward from the lower surface of the body portion 100 to allow a reaction between the processing solution and a reaction reagent, and an outlet portion 33 through which the processing solution flows out from the well portion 32. The reaction reagent that reacts with the processing solution may be fixedly accommodated in the well portion 32. According to the present embodiment, the test chamber 30 is formed substantially similar to the reaction chambers 20.

[0062] The processing-solution test chamber portion 300 may include at least one test chamber 30. When the processing-solution test chamber portion 300 includes a plurality of test chambers 30, the processing-solution test chamber portion 300 may include a flow path through which the processing solution is introduced, and the test chambers 30 may include a reaction reagent for detecting whether a specific target genetic material is included in the processing solution. In the embodiment shown in FIG. 2, four test chambers 30 are provided, and a processing-solution flow path 50 may connect the four test chambers 30 to each other. According to another embodiment as illustrated in FIG. 6, one test chamber 30 may be provided.

[0063] Referring to FIG. 2, a plurality of test chambers 30 are provided in the body portion 100. The test chambers 30 are connected to each other through the processing-solution flow path 50 such that the processing solution may flow in one direction. That is, the processing-solution flow path 50 and the test chambers 30 are connected in series to each other. The processing-solution inlets 12 through which the processing solution may be introduced are connected to both ends of the test chambers 30 that are connected in series to each other. A user may introduce a processing solution to be tested using any one of the processing-solution inlets 12.

[0064] According to the present embodiment, when a plurality of test chambers 30 are provided, the processing-solution flow path 50 for connecting the test chambers 30 to each other includes at least one curved portion in a section connecting one test chamber 30 and an adjacent test chamber 30. The processing-solution flow path 50 is preferably formed to have a size of less than 0.55 mm to prevent excessive pressure during movement of a processing solution and thus ensure smooth movement thereof. Each of the test chambers 30 is provided in the form of a groove extending in one direction.

[0065] The inlet portion 31 of each of the test chambers 30 includes an expanded portion 311 that gradually widens in a direction toward the test chamber 30. The expanded portion 311 is tapered such that when the processing solution flows into the test chamber 30, the processing solution is decelerated by a widening section and fills the internal space of the test chamber 30. The outlet portion 33 of each of the test chambers 30 includes a reduced portion 331 that gradually narrows in a direction away from the test chamber 30 for a processing solution flowing out from the test chamber 30. Due to the reduced portion 331, the flow speed of a processing solution increases, allowing the processing solution to easily move to an adjacent test chamber 30.

[0066] A bottom surface of the test chamber 30 is deeper than a bottom surface of the processing-solution flow path 50. The inlet portion 31 includes an inclined surface 34 connected to the bottom surface of the test chamber 30, and the outlet portion 33 similarly includes an inclined surface 34 connected to the bottom surface of the test chamber 30. The bottom surface of the test chamber 30 is deeper than the bottom surface of the processing-solution flow path 50, thereby forming sufficient space for fixing a reaction reagent, and because the reaction reagent is fixed to the deeper bottom surface, the reaction agent may have improved reactivity with a processing solution. When a fluorescent substance is used, the fluorescent substance may be more easily checked because the well portion 32 is deepened.

[0067] According to the present embodiment, the processing-solution flow path 50 is provided to connect a plurality of test chambers 30 to each other. When the test chambers 30 are referred to as N-th chambers (where N is a natural number) in an order adjacent to a processing-solution inlet 12, the processing-solution flow path 50 connects an N-th chamber and an (N+1)-th chamber. Here, the expression "in the order adjacent to a processing-solution inlet 12" refers to an order in which a processing solution passes through the test chambers 30 sequentially through the processing-solution flow path 50, starting from a reaction chamber 20 that the processing solution first reaches after being injected through the processing-solution inlet 12. According to the present embodiment, the processing-solution flow path 50 includes a third flow path 51, a fourth flow path 52, and a second connection flow path 53.

[0068] The third flow path 51 has one end connected to the N-th chamber and the other end extending toward the (N+1)-th chamber. The fourth flow path 52 has one end connected to the (N+1)-th chamber and the other end extending toward the N-th chamber. The second connection flow path 53 is provided between the third and fourth flow paths 51 and 52 to connect the third and fourth flow paths 51 and 52 to each other. That is, one end of the second connection flow path 53 is connected to the other end of the third flow path 51, and the other end of the second connection flow path 53 is connected to the end of the fourth flow path 52. According to the present embodiment, the third and fourth flow paths 51 and 52 include a bending portion 54 for changing the flow direction of a processing solution. At least one bending portion 54 may be provided.

[0069] According to the present embodiment, as illustrated in FIG. 3, the processing-solution flow path 50 is designed such that a processing solution changes direction three times while moving from the third flow path 51 to the second connection flow path 53, and three times while moving from the second connection flow path 53 to the fourth flow path 52. However, the number of bending portions 54 is not limited thereto and may vary. The bending portions 54 prevent the processing solution from moving between adjacent test chambers 30 when the processing solution reacts with reagents after being completely injected into the test chambers 30 provided in the body portion 100, and thus, reaction results of each test chamber 30 may be reliably secured without being mixed with reaction results of other test chambers 30. The test chambers 30 may include a plurality of reaction reagents for detecting specific target genetic materials.

[0070] According to the present embodiment, a cover portion 60 is coupled to the lower surface of the body portion 100.

[0071] The cover portion 60 is provided to close the sample test chamber portion 200 and the processing-solution test chamber portion 300, which are provided in the form of grooves recessed upward from the lower surface of the body portion 100. When the cover portion 60 is attached to the bottom surface of the body portion 100 and the body portion 100 is positioned such that the cover portion 60 faces downward, the cover portion 60 forms bottom surfaces of the sample test chamber portion 200 and the processing-solution test chamber portion 300. That is, when a sample and a processing solution injected through the sample inlets 11 and the processing-solution inlets 12 are delivered to the sample test chamber portion 200 and the processing-solution test chamber portion 300, bottom surfaces of the sample flow path 40 and the processing-solution flow path 50 are formed by the cover portion 60.

[0072] According to the present embodiment, the cover portion 60 is provided in the form of a film disposed on a lower side of the body portion 100 and attached to the bottom surface of the body portion 100, and heat for performing PCR may be transferred through the cover portion 60 from a heating device. The cover portion 60 may include a thin film, a metal having high heat transfer efficiency, or a polymer material containing nano-carbon to easily receive heat from the heating device. In addition, a sample and a processing solution may automatically move in the sample flow path 40 and the processing-solution flow path 50 without using separate power. An adhesive region R is provided on the bottom surface of the body portion 100 such that the cover portion 60 provided in the form of a film may be attached to the adhesive region R. The adhesive region R is wider than the sample flow path 40 and the processing-solution flow path 50, thereby preventing leakage when a processing solution or a sample is injected. The adhesive region R is formed at edges of the reaction chambers 20, the test chambers 30, the sample flow path 40, and the processing-solution flow path 50, and at an edge of the body portion 100. The adhesive region R may have a width of 0.8 mm to 1.0 mm. When the width of the adhesive region R is less than 0.8 mm, adhesion of the cover portion 60 decreases, and when the width exceeds 1.0 mm, a space for arrangement of the reaction chambers 20 is limited.

[0073] In addition, multi-functional PCR test chips 1001, 1002, and 1003 according to embodiments of the present disclosure are inserted into PCR equipment to perform PCR. The PCR equipment may include: a main body in which the chips 1001, 1002, and 1003 of the present disclosure are mounted; a heating unit provided inside the main body to provide heat to the chips 1001, 1002, and 1003; a light source for projecting light to the chips 1001, 1002, and 1003; a camera for capturing images in response to fluorescence generated when a reaction reagent and a sample react with each other; and an analysis unit for analyzing the images captured using the camera. The present disclosure relates to the chips 1001, 1002, and 1003, and thus a detailed description of the PCR equipment is omitted.(1) First embodiment

[0074] According to an embodiment of the present disclosure, a plurality of reaction chambers 20 and a plurality of test chambers 30 may be provided. For example, the plurality of reaction chambers 20 may be arranged adjacent to one another, and at least one test chamber 30 may be provided on each of both outermost sides of the plurality of reaction chambers 20. For example, in the multi-functional PCR test chip 1001 illustrated in FIG. 4, the sample test chamber portion 200 may include six reaction chambers 20, and the processing-solution test chamber portion 300 may include four test chambers 30. According to the present embodiment, one of the test chambers 30 is adjacent to one side of the six reaction chambers 20 arranged side by side, and the remaining three test chambers 30 are arranged side by side adjacent to an endmost one of the reaction chambers 20. In this case, the three test chambers 30 may be arranged adjacent to one another.

[0075] Specifically, the chip according to the embodiment shown in FIG. 4 may be utilized as follows. Reaction reagents capable of detecting six target genes (hereinafter referred to as target genes 1 to 6) are inserted together into a first test chamber 30, and reaction reagents each capable of detecting one target gene are inserted in second to seven reaction chambers 20, respectively.

[0076] That is, the reaction reagent capable of detecting the target gene 1 may be fixed to a second reaction chamber 20, the reaction reagent capable of detecting the target gene 2 may be fixed to a third reaction chamber 20, the reaction reagent capable of detecting the target gene 3 may be fixed to a fourth second reaction chamber 20, the reaction reagent capable of detecting the target gene 4 may be fixed to a fifth second reaction chamber 20, the reaction reagent capable of detecting the target gene 5 may be fixed to a sixth second reaction chamber 20, and the reaction reagent capable of detecting the target gene 6 may be fixed to a seventh second reaction chamber 20. In addition, the target genes 1 and 2 and the reaction reagents capable of detecting the target genes 1 and 2 may be fixed to an eighth test chamber 30, the target genes 3 and 4 and the reaction reagents capable of detecting the target genes 3 and 4 may be fixed to a ninth test chamber 30, and the target genes 5 and 6 and the reaction reagents capable of detecting the target genes 5 and 6 may be fixed to a tenth test chamber 30.

[0077] The positions of reaction reagents for detecting target genes may be varied depending on conditions, and the numbers of reaction chambers 20 and test chambers 30 may also be varied depending on designs. In this state, a processing solution (lysis buffer) is injected into the first test chamber 30 until the eighth to tenth test chambers 30 are filled with the processing solution, and a sample is injected into the second reaction chamber 20 until the seventh reaction chamber 20 is filled with the sample. Then, PCR is performed.

[0078] No reaction occurs in the first test chamber 30 because the processing solution is not contaminated with the target genes 1 to 6, verifying that the processing solution is not contaminated (NC). In addition, when amplification occurs in the chip inserted into the PCR equipment, successful amplification in the eighth to tenth test chambers 30 may verify that the PCR equipment and / or the reaction reagents function properly.

[0079] In addition, an internal control (IC) may be introduced into the reaction chambers 20 in order to verify whether nucleic acid extraction has been properly performed. The IC is used to verify an inherent gene that must be included in a specimen, and validity of nucleic acid extraction from a sample may be determined by determining, using the IC, whether extracted nucleic acids satisfy a quantitative criterion. The IC may be fixed to any one of the reaction chambers 20, or a user may directly introduce the IC into the reaction chambers 20 before PCR is performed.

[0080] In addition, two types of reaction reagents may be fixed to each reaction chamber to detect two target genetic materials in each reaction chamber. In this case, a total of twelve target genes may be detected in the embodiment shown in FIG. 4 because six reaction chambers 20 are provided. Correspondingly, reaction agents that reacts with 1 to 12 target genetic materials may be fixed to the first test chamber 30 to use the first test chamber 30 as a negative control (NC) for detecting whether the processing solution is contaminated with the twelve target genes. In addition, the eighth to tenth test chambers 30 may be used as a positive control (PC) for verifying the PCR equipment and the reagents.

[0081] As described above, PCR equipment, reaction reagents, and / or PCR processes may be verified using the test chambers 30 and the reaction chambers 20 In the state described above, when a processing solution or DW is introduced into the test chambers 30, no reaction with target genes accommodated in the test chambers 30 may verify that the processing solution is not contaminated (negative control: NC), smooth PCR amplification may verify that reaction agents function properly (positive control: PC).

[0082] In addition, a sample may be introduced into the reaction chambers 20 to test whether target genes are detected. That is, contamination of a processing solution, proper functioning of reagents, and testing of a sample may be performed using a single chip by injecting the processing solution into the test chambers 30 and the sample into the reaction chambers 20.(2) Second embodiment

[0083] FIG. 6 illustrates a multi-functional PCR test chip 1002 according to another embodiment of the present disclosure. As illustrated in FIG. 6, the present embodiment differs from the embodiment shown in FIG. 4 in the positions of sample inlets 11 and processing-solution inlets 12 on an upper side of a body portion 100. As illustrated in FIG. 7, the multi-functional PCR test chip 1002 of the present embodiment includes a plurality of reaction chambers 20 and one test chamber 30. Specifically, as illustrated in FIG. 8, a sample test chamber portion 200 and a processing-solution test chamber portion 300 are provided in a bottom surface of the body portion 100. The sample test chamber portion 200 includes nine reaction chambers 20, and the processing-solution test chamber portion 300 includes one test chamber 30.

[0084] Two types of reaction reagents for detecting two target genetic materials may be fixed to each of the reaction chambers 20. That is, in this case, a total of eighteen target genes may be detected. Correspondingly, reaction reagents capable of reacting with the eighteen target genes may be fixed to the test chamber 30 to detect whether a processing solution is contaminated with the eighteen target genes. Alternatively, a reaction reagent capable of detecting one target genetic material may be fixed to each of the reaction chambers 20.

[0085] In such a state, when no reaction is detected between the eighteen target genes and a processing solution or DW injected into the test chamber 30, the processing solution may be determined not to be contaminated, thereby enabling negative control (NC) testing. In addition, a sample may be injected into the reaction chambers 20 to test whether the target genes are detected. That is, when a test is performed by injecting a sample into the reaction chambers 20, contamination of a processing solution may be checked in the same chip.

[0086] In addition, an internal control (IC) may be injected into any one of the reaction chambers 20 verify whether nucleic acid extraction has been properly performed. The IC is a primer or probe capable of detecting an inherent gene possessed by a living organism, and may be used to check whether nucleic acid extraction from a specimen (sample) has been properly performed. Using the IC, the validity of nucleic acid extraction from the sample may be determined by assessing whether the extracted nucleic acids satisfy a quantitative criterion. The IC may be fixed to any one of the reaction chambers 20, or a user may directly inject the IC into the reaction chambers 20 before PCR is performed.

[0087] In the present embodiment, when a processing-solution test chamber portion 200 is configured by connecting a first chamber and a tenth chamber to each other, and a sample test chamber portion 200 is configured by connecting section to nineth chambers to each other, the tenth chamber may be used to function as a positive control (PC) as in the first embodiment.(3) Third embodiment

[0088] According to another embodiment of the present disclosure, a multi-functional PCR test chip 1003 may include a plurality of sample test chamber portions 200 and a plurality of processing-solution test chamber portions 300. When PCR is performed, any one of a plurality of sample test chamber portions 201 and 202 and any one of a plurality of processing-solution test chamber portions 301 and 302 may be used together as a set. That is, according to the present embodiment, a plurality of sets each including a combination of the sample test chamber portions 201 and 202 and the processing-solution test chamber portions 301 and 302 may be provided.

[0089] Referring to FIG. 9, the sample test chamber portions 200 include a first sample test chamber portion 201 including a plurality of reaction chambers 20 provided adjacent to one another, and a second sample test chamber portion 202 separated from the first sample test chamber portion 201 and including another plurality of reaction chambers 20 provided adjacent to one another. In addition, the processing-solution test chamber portion 300 may include at least one first test chamber 301 adjacent to the first sample test chamber portion 201, and at least one second test chamber 302 adjacent to the second sample test chamber portion 202. According to the present embodiment, the first sample test chamber portion 201 and the second sample test chamber portion 202 are disposed adjacent to each other between the first test chamber 301 and the second test chamber 302.

[0090] According to the present embodiment, as illustrated in FIG. 9, the first test chamber 301 and the first sample test chamber portion 201 form one set, and the second sample test chamber portion 202 and the second test chamber 302 form another set.

[0091] Specifically, the first sample test chamber portion 201 includes four reaction chambers 20 arranged adjacent to one another and connected to each other through a sample flow path 40. The second sample test chamber portion 202 is not connected to the first sample test chamber portion 202, and includes four reaction chambers 20 arranged adjacent to one another. The second sample test chamber portion 202 also includes a sample flow path 40 as a connection path.

[0092] The number of reaction chambers 20 of each of the first and second sample test chamber portions 201 and 202 is not limited to the four described above and may vary. Referring to FIG. 9, the first test chamber 301 is disposed adjacent to a left outermost side of the first sample test chamber portion 201, and the second test chamber 302 is disposed adjacent to a right outermost side of the second sample test chamber portion 202. The first test chamber 301 and the first sample test chamber portion 201 are formed symmetrically with the second test chamber 302 and the second sample test chamber portion 202. Alternatively, the first test chamber 301 may be provided on a right outermost side of the first sample test chamber portion 201, or the second test chamber 302 may be provided on a left outermost side of the second sample test chamber portion 202.

[0093] Two processing-solution inlets 12 are provided in each of the first and second test chambers 301 and 302. Two sample inlets 11 are provided at both ends of each of the first sample test chamber portion 201 and the second sample test chamber portion 202. According to the present embodiment, the first test chamber 301 may be used to test contamination of a processing solution or DW used to process a first sample, and the first sample test chamber portion 201 may be used to test whether a target gene is detected in the first sample introduced into the reaction chambers 20. In addition, the second test chamber 302 may be used to test contamination of a processing solution or DW used to process a second sample, and the second sample test chamber portion 202 may be used to test whether a target gene is detected in the second sample introduced into the reaction chambers 20.

[0094] Referring again to FIG. 9, two types of reaction reagents capable of detecting two target genetic materials may be fixed to each of second to fifth reaction chambers 20 of the first sample test chamber portion 201. That is, in this case, a total of eight target genes may be detected. Correspondingly, reaction reagents capable of reacting with the eight target genetic materials may be fixed to the first test chamber 301 to detect whether the processing solution for processing the first sample is contaminated with the eight target genes. Alternatively, a reaction reagent for detecting one target genetic material may be fixed to each of the reaction chambers 20 of the first sample test chamber portion 201.

[0095] In such a state, when the processing solution or DW for processing the first sample is introduced into the first test chamber 301, and no reaction is detected between the processing solution or DW and the eight target genes, the processing solution is determined not to be contaminated such that the first test chamber may function as a negative control (NC). In addition, the first sample may be introduced into the first sample test chamber portion 201 to test whether target genes are detected.

[0096] That is, when a test is performed by introducing the first sample into the reaction chambers 20, contamination of the processing solution for processing the first sample may be checked in the same chip. In addition, an internal control (IC) may be introduced into any one of the reaction chambers 20 to verify whether nucleic acid extraction has been properly performed. As described above, using the IC, validity of nucleic acid extraction from a sample may be determined by assessing whether the extracted nucleic acids satisfy a quantitative criterion. The IC may be fixed to any one of the reaction chambers 20 of the first sample test chamber portion 201, or a user may directly introduce the IC into the reaction chambers 20 before PCR is performed.

[0097] Alternatively, in the present embodiment, first and fifth chambers may be connected to each other to form the first test chamber 301, and second to fourth chambers may be connected to each other to form the first sample test chamber portion 201. In this case, the fifth chamber may be used to function as a positive control (PC) as in the first embodiment.

[0098] Similarly to the first sample test chamber portion 201, in the second sample test chamber portion 202, two kinds of reaction reagents for detecting two target genetic materials may be fixed to each of sixth to nineth reaction chambers 20. That is, in this case, a total of eight target genes may be detected. The eight target genes may be the same as the eight target genes of the first sample test chamber portion 201. reaction reagents capable of reacting with the eight target genetic materials may be fixed to the second test chamber 302 to detect whether the processing solution for processing the second sample is contaminated with the eight target genes, Alternatively, a reaction reagent for detecting one target genetic material may be fixed to each of the reaction chambers 20 of the second sample test chamber portion 202.

[0099] In such a state, when the processing solution or DW for processing the second sample is introduced into the second test chamber 302, and no reaction is detected between the processing solution or DW and the eight target genes, the processing solution is determined not to be contaminated such that the first test chamber may function as a negative control (NC). In addition, the second sample may be introduced into the second sample test chamber portion 202 to test whether target genes are detected. That is, when a test is performed by introducing the second sample into the reaction chambers 20, contamination of the processing solution for processing the second sample may be checked in the same chip.

[0100] In addition, an internal control (IC) may be introduced into any one of the reaction chambers 20 to verify whether nucleic acid extraction has been properly performed. As described above, using the IC, validity of nucleic acid extraction from a sample may be determined by assessing whether the extracted nucleic acids satisfy a quantitative criterion. The IC may be fixed to any one of the reaction chambers 20 of the second sample test chamber portion 202, or a user may directly introduce the IC into the reaction chambers 20 before PCR is performed.

[0101] Alternatively, in the present embodiment, sixth and tenth chambers may be connected to each other to form the second test chamber 302, and seventh to nineth chambers may be connected to each other to form the second sample test chamber portion 202. In this case, the sixth chamber may be used to function as a positive control (PC) as in the first embodiment.

[0102] As described above, the present embodiment provides a chip capable of simultaneously testing different samples. According to the present embodiment, two samples may be tested with one chip, and in this case, contamination of processing solutions for processing the two samples may be simultaneously checked. The numbers of reaction chambers and test chambers or the types and numbers of reaction reagents fixed to the reaction chambers and the test chambers may be varied depending on target genetic materials to be detected.

[0103] Through the processes described above, the present disclosure enables multiplex testing by detecting multiple types of target genes with a single injection of a sample, and also enables contamination of a processing solution, such as a buffer used to extract nucleic acids from the sample, to be checked together on a single chip. That is, a sample and a processing solution may be simultaneously tested owing to a configuration in which a sample test chamber portion, including a plurality of reaction chambers into which the sample is introduced, is separated from a processing-solution test chamber portion that functions as a negative control (NC) by receiving the processing solution or the like to check whether the processing solution is contaminated.

[0104] In addition, because the processing-solution test chamber portion includes a plurality of test chambers for checking whether reaction reagents function properly (positive control (PC)), testing of a sample, testing of contamination of a processing solution, and checking whether reaction reagents function properly may be simultaneously performed.

[0105] In addition, the present disclosure aims to enable simultaneous multiplex diagnosis with high sensitivity and specificity. In multiplex PCR diagnosis of the related art, a plurality of primer probes for detecting multiple targets are disposed in the same space, leading to interference, competitive reactions, and variations in reaction characteristics, thereby degrading sensitivity and specificity. To address this problem, target detection regions are spatially separated from each other to eliminate mutual interference and competitive reactions. In addition, because reaction reagents react with a sample in a state in which the reaction agents are fixed to reaction chambers, sensitivity may be improved when checking test results.

[0106] While preferred embodiments of the present disclosure have been described above in detail, the present disclosure is not limited to the embodiments, and various modifications may be made without departing from the scope of the present disclosure.

Examples

first embodiment

(1) First embodiment

[0074]According to an embodiment of the present disclosure, a plurality of reaction chambers 20 and a plurality of test chambers 30 may be provided. For example, the plurality of reaction chambers 20 may be arranged adjacent to one another, and at least one test chamber 30 may be provided on each of both outermost sides of the plurality of reaction chambers 20. For example, in the multi-functional PCR test chip 1001 illustrated in FIG. 4, the sample test chamber portion 200 may include six reaction chambers 20, and the processing-solution test chamber portion 300 may include four test chambers 30. According to the present embodiment, one of the test chambers 30 is adjacent to one side of the six reaction chambers 20 arranged side by side, and the remaining three test chambers 30 are arranged side by side adjacent to an endmost one of the reaction chambers 20. In this case, the three test chambers 30 may be arranged adjacent to one another.

[0075]Specifically, the ...

second embodiment

(2) Second embodiment

[0083]FIG. 6 illustrates a multi-functional PCR test chip 1002 according to another embodiment of the present disclosure. As illustrated in FIG. 6, the present embodiment differs from the embodiment shown in FIG. 4 in the positions of sample inlets 11 and processing-solution inlets 12 on an upper side of a body portion 100. As illustrated in FIG. 7, the multi-functional PCR test chip 1002 of the present embodiment includes a plurality of reaction chambers 20 and one test chamber 30. Specifically, as illustrated in FIG. 8, a sample test chamber portion 200 and a processing-solution test chamber portion 300 are provided in a bottom surface of the body portion 100. The sample test chamber portion 200 includes nine reaction chambers 20, and the processing-solution test chamber portion 300 includes one test chamber 30.

[0084]Two types of reaction reagents for detecting two target genetic materials may be fixed to each of the reaction chambers 20. That is, in this case...

third embodiment

(3) Third embodiment

[0088]According to another embodiment of the present disclosure, a multi-functional PCR test chip 1003 may include a plurality of sample test chamber portions 200 and a plurality of processing-solution test chamber portions 300. When PCR is performed, any one of a plurality of sample test chamber portions 201 and 202 and any one of a plurality of processing-solution test chamber portions 301 and 302 may be used together as a set. That is, according to the present embodiment, a plurality of sets each including a combination of the sample test chamber portions 201 and 202 and the processing-solution test chamber portions 301 and 302 may be provided.

[0089]Referring to FIG. 9, the sample test chamber portions 200 include a first sample test chamber portion 201 including a plurality of reaction chambers 20 provided adjacent to one another, and a second sample test chamber portion 202 separated from the first sample test chamber portion 201 and including another plural...

Claims

1. A multi-functional PCR test chip comprising: a body portion comprising a light-transmissive material, a sample inlet through which a sample is introduced, and a processing-solution inlet through which a processing solution for processing the sample is introduced; a sample test chamber portion provided in the body portion and comprising a reaction chamber, the reaction chamber accommodating a reaction reagent for detecting a target genetic material and in which the sample introduced through the sample inlet reacts with the reaction reagent; and a processing-solution test chamber portion provided in the body portion and comprising a test chamber, the test chamber accommodating the reaction reagent and in which the processing solution introduced through the processing-solution inlet reacts with the reaction reagent.

2. The multi-functional PCR test chip of claim 1, wherein the reaction chamber comprises: an inlet portion through which the sample flows in; a reaction well portion in which the sample reacts with the reaction reagent, the reaction well portion being recessed upward from a lower surface of the body portion; and an outlet portion through which the sample flows out from the reaction well portion.

3. The multi-functional PCR test chip of claim 2, wherein the inlet portion and the outlet portion of the reaction chamber are inclined.

4. The multi-functional PCR test chip of claim 2, wherein a plurality of reaction chambers are provided in the body portion, and the plurality of reaction chambers are connected to each other through a sample flow path to allow the sample to flow in one direction.

5. The multi-functional PCR test chip of claim 2, wherein the inlet portion comprises an expanded portion that widens in a direction in which the sample enters the reaction well portion, and the outlet portion comprises a reduced portion that narrows in a direction in which the sample flows out from the reaction well portion.

6. The multi-functional PCR test chip of claim 4, wherein, when the plurality of reaction chambers are referred to as N chambers (where N is a natural number) in order adjacent to the sample inlet, the sample flow path connects an N-th chamber and an (N+1)-th chamber to each other, wherein the sample flow path comprises: a first flow path having one end connected to the N-th chamber; a second flow path having another end connected to the (N+1)-th chamber; and a first connection flow path provided between the first flow path and the second flow path and connecting the first flow path and the second flow path to each other, wherein the first flow path and the second flow path comprise at least one bending portion to change a flow direction of the sample.

7. The multi-functional PCR test chip of claim 1, wherein a cover portion is coupled to a lower surface of the body portion to close the sample test chamber portion and the processing-solution test chamber portion that are provided as grooves recessed upward from the lower surface of the body portion, and in a state in which the body portion is positioned with the cover portion facing downward, the cover portion forms bottom surfaces of the sample test chamber portion and the processing-solution test chamber portion.

8. The multi-functional PCR test chip of claim 7, wherein the cover portion is provided as a film disposed on a lower side of the body portion and attached to the lower surface of the body portion to transfer heat from a heating device for performing PCR.

9. The multi-functional PCR test chip of claim 1, wherein the test chamber comprises a plurality of test chambers.

10. The multi-functional PCR test chip of claim 9, wherein the plurality of test chambers are connected to each other through a processing-solution flow path to allow the processing solution to flow in one direction.

11. The multi-functional PCR test chip of claim 10, wherein, when the plurality of test chambers are referred to as M chambers (where M is a natural number) in order adjacent to the processing-solution inlet, the processing-solution flow path connects an M-th chamber and an (M+1)-th chamber to each other, wherein the processing-solution flow path comprises: a third flow path having one end connected to the M-th chamber; a fourth flow path having another end connected to the (M+1)-th chamber; and a second connection flow path provided between the third flow path and the fourth flow path and connecting the third flow path and the fourth flow path to each other, wherein the third flow path and the fourth flow path comprise at least one bending portion to change a flow direction of the sample.

12. The multi-functional PCR test chip of claim 4, wherein the plurality of reaction chambers are arranged adjacent to one another, and the test chamber comprises a plurality of test chambers, at least one of the plurality of test chambers being provided on each of both outermost sides of the plurality of reaction chambers arranged adjacent to one another.

13. The multi-functional PCR test chip of claim 1, wherein the sample test chamber portion comprises: a first sample test chamber portion comprising a plurality of reaction chambers arranged adjacent to each other; and a second sample test chamber portion provided apart from the first sample test chamber portion and comprising another plurality of reaction chambers arranged adjacent to each other, wherein the processing-solution test chamber portion comprises at least one first test chamber adjacent to the first sample test chamber portion and at least one second test chamber adjacent to the second sample test chamber portion.

14. The multi-functional PCR test chip of claim 13, wherein the first sample test chamber portion and the second sample test chamber portion are arranged adjacent to each other between the first test chamber and the second test chamber.

15. The multi-functional PCR test chip of claim 1, wherein reaction reagents for detecting at least two different target genes are fixed to the reaction chamber and the test chamber to enable multiplex diagnosis for samples.

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