Multi-functional PCR test chip

Abstract

The present invention is characterized by comprising: a body part made of a light-transmitting material and having a sample inlet through which a sample is introduced and a processing liquid inlet through which a processing liquid for processing the sample is introduced; a sample test chamber part, provided in the body part and comprising a reaction chamber which accommodates a reaction reagent for detecting a target genetic material and in which a sample injected through the sample inlet and the reaction reagent react with each other; and a processing liquid test chamber part, provided in the body part and comprising a test chamber which accommodates the reaction reagent and in which the processing liquid injected through the processing liquid inlet and the reaction reagent react.

Description

Multifunctional PCR test chip

[0001] The present invention relates to a multifunctional PCR test chip, and relates to a chip that reacts a sample with a reaction reagent to perform a PCR test. The reaction reagent is fixed to the chip, and the chip is used to perform a PCR test by introducing a sample to be tested into the chip and checking whether the sample reacts with the reaction reagent.

[0002] Polymerase Chain Reaction (PCR) is a method for amplifying specific target genetic material. It is a technology that amplifies a small amount of genetic material with identical base sequences into a large quantity. PCR is used to diagnose various genetic diseases by amplifying human nucleic acids, and is also applied to bacterial, viral, and fungal nucleic acids for the diagnosis of infectious diseases.

[0003] In general, PCR is performed repeatedly through the following three steps. The three steps include: 1) a denaturing step in which a sample solution containing double-stranded DNA is heated to a specific temperature, for example, about 95°C, to separate the double-stranded DNA into single-stranded DNA; 2) an annealing step in which, after the denaturing step, an oligonucleotide primer having a sequence complementary to a specific base sequence to be amplified is provided to the sample solution, and the solution is cooled to a specific temperature, for example, 55°C, together with the separated single-stranded DNA, to bind the primer to the specific base sequence of the single-stranded DNA to form a partial DNA-primer complex; and 3) an extension (or amplification) step in which, after the annealing step, the sample solution is maintained at an active temperature of a DNA polymerase, for example, 72°C, to form double-stranded DNA based on the primer of the partial DNA-primer complex by a DNA polymerase. By repeating the above three steps several times, a target nucleic acid having a specific base sequence can be exponentially amplified. However, in the case of RNA, a reverse transcription step into DNA may be performed prior to steps 1) to 3).

[0004] Performing this type of PCR requires a PCR chip equipped with a chamber capable of holding reagents and samples, and a PCR device that heats and cools the PCR chip to induce an amplification reaction and measure the results. PCR devices are a specific means of amplifying target nucleic acids, and various devices have been developed.

[0005] Conventional PCR could only qualitatively analyze the amplified target nucleic acid using separate electrophoresis after the reaction was completed. However, recently, real-time PCR devices (i.e., rPCR devices) have been developed that can quantitatively analyze the target nucleic acid by detecting the intensity of fluorescence proportional to the concentration of the amplified genetic material using an optical detection system. In addition, portable real-time PCR devices have been developed that can perform PCR on-site where samples are collected by miniaturizing large PCR devices used in laboratories. Portable PCR devices are characterized by using PCR chips (or biochips, microchips) and using cameras or digital cameras instead of large and complex optical detection systems.

[0006] The present invention relates to a PCR test chip that improves sensitivity when detecting the intensity of fluorescence emitted when a reagent and a sample react, enables simultaneous detection of multiple target genes, and also enables checking for contamination of a processing solution such as a buffer used to extract nucleic acids from a sample.

[0007] Conventional techniques utilize a processing solution (buffer solution) to extract nucleic acids from a sample, then inject the nucleic acids contained in the buffer solution into a chip to test for interaction with the reaction reagent. However, this method requires additional work to confirm contamination of the processing solution, which is inconvenient.

[0008] The present invention has been devised to solve the above-mentioned problem, and the purpose of the present invention is to provide a PCR test chip in which a reaction reagent that reacts with a sample is accommodated and fixed in a reaction chamber so as to improve sensitivity when confirming a test result, and also to check for contamination of a treatment solution such as a buffer used to extract nucleic acid from a sample.

[0009] A multifunctional PCR test chip according to one embodiment of the present invention is characterized by including: a body part formed of a light-transmitting material and having a sample inlet for introducing a sample and a treatment solution inlet for introducing a treatment solution for processing the sample; a sample test chamber part provided in the body part and including a reaction chamber in which a reaction reagent for detecting a target genetic material is accommodated and in which a sample introduced through the sample inlet reacts with the reaction reagent; and a treatment solution test chamber part provided in the body part and including a test chamber in which the reaction reagent is accommodated and in which a treatment solution introduced through the treatment solution inlet reacts with the reaction reagent.

[0010] In addition, it is preferable that the reaction chamber includes an inlet portion into which the sample is introduced; a reaction well portion formed by sinking from the lower surface of the body portion toward the upper surface, where the sample reacts with the reaction reagent; and an outlet portion through which the sample flows out from the reaction well portion.

[0011] In addition, it is preferable that the inlet and outlet of the reaction chamber are formed to be inclined.

[0012] In addition, it is preferable that a plurality of reaction chambers are provided in the body portion, and that the reaction chambers are connected to each other by a sample movement path so that the sample flows in one direction.

[0013] In addition, it is preferable that the inlet portion includes an expansion portion whose width increases in the direction in which the sample enters the reaction well portion, and that the outlet portion includes a reduction portion whose width decreases in the direction in which the sample flows out of the reaction well portion.

[0014] In addition, the sample movement path, when the reaction chamber is referred to as the Nth chamber (N is a natural number) in the order adjacent to the sample inlet, connects the Nth chamber and the N+1th chamber, and the sample movement path includes a first path having one end connected to the Nth chamber; a second path having the other end connected to the N+1th chamber; and a first connecting path connecting the first path and the second path; and it is preferable that the first path and the second path include at least one bending portion so that the flow direction of the sample is changed.

[0015] In addition, on the lower surface of the body part, a cover part that closes the chamber part for sample inspection and the chamber part for treatment liquid inspection, which are provided in a groove shape from the lower surface toward the upper surface, are combined, and it is preferable that the cover part forms the bottom surface of the chamber part for sample inspection and the chamber part for treatment liquid inspection when the body part is positioned so that the cover part faces downward.

[0016] In addition, the cover part is preferably provided in the form of a film and attached to the lower surface of the body part so as to be positioned on the lower side of the body part and to transmit heat from a heating device for performing PCR.

[0017] In addition, it is preferable that a plurality of the above inspection chambers are provided.

[0018] In addition, it is preferable that the inspection chambers are connected to each other by a treatment liquid flow path so that the treatment liquid flows in one direction.

[0019] In addition, the treatment liquid movement path, when the inspection chamber is referred to as the M chamber (M is a natural number) in the order adjacent to the treatment liquid injection port, connects the M chamber and the M+1 chamber, and the treatment liquid movement path includes a third path having one end connected to the M chamber; a fourth path having the other end connected to the M+1 chamber; and a second connecting path connecting the third path and the fourth path; and it is preferable that the third path and the fourth path include at least one bending portion so that the flow direction of the sample is changed.

[0020] In addition, the plurality of reaction chambers are arranged adjacent to each other, and the inspection chambers are provided in plurality, and it is preferable that at least one inspection chamber is provided on both outermost sides of the plurality of reaction chambers arranged adjacent to each other.

[0021] In addition, it is preferable that the sample inspection chamber section includes a first sample inspection chamber section in which a plurality of reaction chambers are provided adjacent to each other, and a second sample inspection chamber section in which a plurality of other reaction chambers are provided adjacent to each other and separated from the first sample inspection chamber section, and the treatment liquid inspection chamber section includes at least one first inspection chamber adjacent to the first sample inspection chamber section and at least one second inspection chamber adjacent to the second sample inspection chamber section.

[0022] In addition, the first sample inspection chamber part and the second sample inspection chamber part are preferably arranged adjacent to each other, but are preferably arranged between the first inspection chamber and the second inspection chamber.

[0023] In addition, it is preferable that reaction reagents for detecting at least two different target genes are fixed in the reaction chamber and the inspection chamber, so that multiple diagnosis is possible for each sample.

[0024] A multifunctional PCR test chip according to an embodiment of the present invention provides the effect of improving sensitivity when confirming test results by allowing a reaction reagent that reacts with a sample to be accommodated and fixed within a reaction chamber, and also enabling confirmation of contamination of a processing solution, such as a buffer used to extract nucleic acid from a sample.

[0025] Specifically, the present invention was developed for the purpose of simultaneous multiplex diagnosis with high sensitivity and specificity. In order to solve the problem that in the existing multiplex PCR diagnosis, a large number of primer probes that detect multiple targets are gathered in the same space, which leads to a decrease in sensitivity and specificity due to interference and competitive reactions and different reaction characteristics among them, the detection zones of each target are spatially separated, so that interference and competitive reactions among them can be eliminated.

[0026] In addition, unlike existing multiplex PCR diagnostics, there is no need to prepare multiple reaction solutions, and since it is connected to a single path, the reaction can be carried out simply and quickly with a single reaction solution preparation and injection.

[0027] Additionally, although connected by a single channel, it is designed to prevent diffusion or interference between each target space by applying microfluidic technology, which can improve the accuracy of test results.

[0028] In addition, separate paths are designed for contamination (NC: negative control) and equipment and reagent verification (PC: positive control), providing the effect of performing NC and / or PC together as well as sample inspection.

[0029] In addition, the sample movement path connecting the reaction chambers and the treatment liquid movement path connecting the inspection chambers are provided with a banding portion so that when the treatment liquid or sample reacts with the reaction reagent, the treatment liquid or sample is effectively prevented from moving from one reaction chamber or inspection chamber to an adjacent reaction chamber or inspection chamber, thereby providing the effect of improving the accuracy of the test results.

[0030] In addition, the reaction chamber or inspection chamber includes a sunken well for reacting with a reaction reagent, so that the reaction is performed in the well, thereby improving visibility, and the reaction chamber or inspection chamber has an expansion portion provided at the inlet portion and a reduction portion provided at the outlet portion, thereby providing the effect of allowing a sample or treatment liquid to be dispensed (spread) quickly.

[0031] Figure 1 is a perspective view of a multifunctional PCR test chip according to one embodiment of the present invention.

[0032] Figure 2 is a bottom perspective view of Figure 1.

[0033] Figure 3 is an enlarged view of Figure 2.

[0034] Figure 4 is a drawing showing the functions of the reaction chamber and inspection chamber of Figure 1.

[0035] Figure 5 is a cross-sectional view of Figure 1;

[0036] Figure 6 is a perspective view of a multifunctional PCR test chip according to another embodiment of the present invention.

[0037] Fig. 7 is a bottom perspective view of Fig. 6.

[0038] Figure 8 is a drawing showing the functions of the reaction chamber and inspection chamber of Figure 1.

[0039] FIG. 9 is a drawing illustrating a multifunctional PCR test chip according to another embodiment of the present invention.

[0040] Hereinafter, various embodiments of the present invention will be described in connection with the accompanying drawings. Various embodiments of the present invention may have various modifications and various embodiments, and thus specific embodiments are illustrated in the drawings and related detailed descriptions are provided. However, this is not intended to limit the various embodiments of the present invention to specific embodiments, but should be understood to include all modifications and / or equivalents or substitutes included in the spirit and technical scope of the various embodiments of the present invention. In connection with the description of the drawings, similar reference numerals have been used for similar components.

[0041] Expressions such as “includes” or “may include” that may be used in various embodiments of the present invention indicate the existence of the disclosed function, operation, or component, etc., and do not limit one or more additional functions, operations, or components, etc. In addition, in various embodiments of the present invention, it should be understood that terms such as “includes” or “has” are intended to specify the existence of a feature, number, step, operation, component, part, or combination thereof described in the specification, and do not exclude in advance the possibility of the existence or addition of one or more other features, numbers, steps, operations, components, parts, or combinations thereof.

[0042] When a component is referred to as being "connected" to another component, it should be understood that the component may be directly connected to the other component, but that there may also be a new component between the component and the other component. Conversely, when a component is referred to as being "directly connected" or "directly connected" to another component, it should be understood that no new component exists between the component and the other component.

[0043] The terms used in the various embodiments of the present invention are used solely to describe specific embodiments and are not intended to limit the various embodiments of the present invention. Singular expressions include plural expressions unless the context clearly indicates otherwise.

[0044] Unless otherwise defined, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by one of ordinary skill in the art to which the various embodiments of the present invention belong.

[0045] Terms defined in commonly used dictionaries should be interpreted as having a meaning consistent with their meaning in the context of the relevant technology, and will not be interpreted in an idealized or overly formal sense unless explicitly defined in various embodiments of the present invention.

[0046]

[0047] The present invention relates to a chip used in a PCR device. A PCR chip accommodates a reaction reagent, and after a sample is injected into the chip, the chip is utilized to detect a target gene using a PCR device. The present invention is characterized by a structure in which a portion having a plurality of reaction chambers into which a sample is injected and a chamber portion for testing for contamination by injecting a treatment solution, reaction reagent, etc. to enable NC (negative control) and / or PC (positive control) are formed separately, so that testing of a sample and testing of a treatment solution can be performed simultaneously.

[0048]

[0049] The term "PCR (polymerase chain reaction)" used herein refers to a reaction that amplifies a specific target nucleic acid molecule using a thermostable DNA polymerase. In addition to the DNA polymerase, PCR may also use a reaction mixture containing primers (forward primers, reverse primers), which are oligonucleotides that can specifically hybridize to the target nucleic acid, a deoxynucleotide triphosphate mixture (dNTP mixture), and divalent ions such as Mg2+.

[0050] A 'primer' is used to initiate a PCR reaction, and refers to an oligonucleotide or polynucleotide that hybridizes complementarily to template DNA. Primers for a PCR reaction may be a pair of forward primers (or sense primers) selected from the sense strand of the nucleic acid molecule being amplified in the same direction as the genetic code progression, and reverse primers (or antisense primers) selected from the antisense strand complementary to the sense strand.

[0051] "Sample" refers to the genetic material to be amplified or a biological solution containing such genetic material. "Reaction reagent" is intended to detect the target genetic material and may include primers or other materials. Primers may be a pair of primers, each 15-30 bp long, capable of binding to both ends of a specific region of the target gene. Furthermore, DNA polymerase uses an enzyme that retains its activity even at high temperatures (90°C or higher).

[0052]

[0053] Hereinafter, a preferred embodiment of the present invention will be described in detail with reference to the attached drawings. According to one embodiment of the present invention, the multifunctional PCR test chip (1001) includes a body part (100), a chamber part (200) for sample testing, and a chamber part (300) for treatment solution testing.

[0054]

[0055] The body part (100) is made of a light-transmitting material so that a fluorescent substance can be detected by an optical detection device. As illustrated in FIG. 1, according to the present embodiment, the body part (100) may be made of a transparent polymer material. The body part (100) may be made in a rectangular shape. Of course, the body part (100) may be made in various shapes such as a circle or a square. According to the present embodiment, in the rectangular body part (100), a protruding wall (13) is formed on the long sides facing each other so that the user can easily grip it. A reinforcing protrusion (14) extending in the vertical direction is formed on the inside of the protruding wall (13). The reinforcing protrusion (14) prevents the injection-molded product (100) from bending during injection molding and improves the rigidity of the body part (100). In addition, the reinforcing projection (14) enables chips to be stacked in an up-and-down direction during product production or on-site, and at this time, the reinforcing projection (14) can perform the function of a support to support the upper chip. Meanwhile, a groove is formed on the outer circumference of the protruding wall (13) so that the user can easily grip the body portion (100).

[0056] In addition, as illustrated in FIGS. 1 and 2, a sample inlet (11) for injecting a sample and a treatment solution inlet (12) for injecting a treatment solution for treating the sample are formed in the body part (100). A groove-shaped flow path (40, 50) and a plurality of chambers (20, 30) are formed in the body part (100). A sealing film (60) is attached to the body part (100) to form a flow path through which the sample and treatment solution move together with the flow paths (40, 50) and the chambers (20, 30).

[0057]

[0058] The chamber part (200) for sample inspection is provided in the body part (100) and includes a reaction chamber (20) in which a reaction reagent for detecting a target genetic material is received and in which a sample introduced through the sample inlet (11) reacts with the reaction reagent. According to the present embodiment, the chamber part (200) for sample inspection includes a flow path extending in one direction and a plurality of reaction chambers (20) provided in the flow path.

[0059] The reaction chamber (20), as illustrated in FIG. 2, includes an inlet (21) into which a sample is introduced, a reaction well (22) formed by recessing from the lower surface of the body (100) toward the upper surface, where the sample and a reaction reagent react, and an outlet (23) through which the sample flows out from the reaction well (22). The reaction chamber (20) may include a reaction reagent that reacts with the sample. Specifically, the reaction reagent that reacts with the sample may be accommodated and fixed in the reaction well (22).

[0060] 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 by a sample movement path (40) so that the sample flows in one direction. That is, the sample movement path (40) and the reaction chamber (20) are connected in series. A sample inlet (11) for introducing a sample is connected to both ends of the reaction chambers (20) connected in series. A user can introduce a sample to be inspected using any one of the sample inlets (11).

[0061] The above reaction reagent may include a primer, etc. In addition, the reaction reagent may also include a fluorescent dye. Of course, depending on the PCR method, the fluorescent dye may be injected into the chip together with the sample. The fluorescent dye emits light of a specific wavelength in the DNA extension (or amplification) step. At least two types of the reaction reagent are provided to detect different target genes, and different reaction reagents may be accommodated in the plurality of reaction chambers (20). The plurality of reaction chambers (20) are provided, and different reaction reagents are fixed in the reaction chambers (20) to detect different target genes with a single sample injection, thereby enabling multiplexed diagnosis.

[0062] For example, in one embodiment, six reaction chambers (20) may be provided. By accommodating and fixing reaction reagents for detecting two specific target genes in each reaction chamber (20), a total of 12 target genes can be detected. At this time, each reaction reagent may be fixed to the reaction chamber (20) in a state in which a fluorescent dye is included. Of course, the fluorescent dye may be injected into the chip together with the sample. The fluorescent dyes fixed together to different reaction reagents or injected together with the sample may use the same first dye, or different first and second dyes. The fluorescent dye reacts to light of a specific wavelength, and the first and second dyes may generate fluorescence in response to light of different wavelengths. By detecting the light emitted when amplification is performed by reacting with the target gene in each reaction chamber, it is possible to identify whether a reaction has occurred in the corresponding reaction chamber (20). If the first dye is equally injected into six chambers, light of a single wavelength that reacts to the first dye can be irradiated to enable six types of multiple diagnosis at once, and if the first dye and the second dye are simultaneously injected into each reaction chamber (20), light of multiple wavelengths having two different wavelengths can be irradiated to enable 12 types of multiple diagnosis at once.

[0063] The above reaction reagent is fixed to the reaction chamber (20), and the chip with the reaction reagent fixed thereon can be transported to a location requiring PCR testing and used as needed. The reaction reagent can be fixed to the reaction chamber (20) by a method such as natural drying, vacuum drying, or freeze drying. In addition, the surface of the reaction reagent fixed to the reaction chamber (20) can be coated using a coating agent having waterproof and adhesive properties.

[0064] The coating agent may be fixed to the reagent (primer, probe, premix) using a biopolymer material that does not affect the properties of the reagent or the PCR results. The coating agent may remain solid at room temperature and may have the property of melting at a certain temperature or higher. For example, the coating agent may melt at 45°C or higher. When the chip into which the sample has been injected is heated during the PCR process, the coating agent may melt and the sample and the reaction reagent may react. The coating agent may be composed of any one of wax, agarose, paraffin, collagen, chitosan, gelatin, or a mixture thereof. Of course, the coating agent may be accommodated and fixed in a reaction chamber mixed with the reaction reagent. Even in this case, the reaction reagent accommodated in the reaction chamber may remain solid at room temperature, and when the chip into which the sample has been injected is heated during the PCR process, the coating agent may melt and the sample and the reaction reagent may react.

[0065]

[0066] According to the present embodiment, the path through which the sample is transferred connects the reaction chambers (20) to each other, and the sample transfer path (40) includes at least one curved portion in the section connecting the reaction chambers (20) and the adjacent reaction chambers (20). It is preferable that the sample transfer path (40) be formed to be less than 0.55 mm to prevent excessive pressure from being generated when the sample is moved, thereby inducing smooth movement. The reaction chamber (20) is provided in the form of a groove that is long in one direction.

[0067] The inlet portion (21) of the reaction chamber (20) includes an expansion portion (211) whose width gradually widens as it approaches the reaction chamber (20). The expansion portion (211) is formed to be tapered, so that when the sample flows into the reaction chamber (20), the speed is reduced by the widening section, thereby filling the internal space of the reaction chamber (20). The outlet portion (23) of the reaction chamber (20) includes a narrowing portion (231) whose width narrows as it moves away from the reaction chamber (20) as the sample flows out of the reaction chamber (20). The flow speed of the sample is increased by the narrowing portion (231), thereby allowing the sample to easily move to another adjacent reaction chamber (20).

[0068] The bottom surface of the reaction chamber (20) is formed deeper than the bottom surface of the sample movement path (40). The inlet (21) has an inclined surface (24) connected to the bottom surface of the reaction chamber (20), and the outlet (23) has an inclined surface (24) connected to the bottom surface of the reaction chamber (20) like the inlet (21). The bottom surface of the reaction chamber (20) is formed deeper than the bottom surface of the sample movement path (40) to form sufficient space for fixing the reaction reagent, and the reaction reagent is fixed to the deeper bottom to improve reactivity with the sample. When a fluorescent substance is used, the deep reaction well (22) provides an effect that makes it easier to identify the fluorescent substance.

[0069] According to the present embodiment, the sample movement path (40) is provided to connect the plurality of reaction chambers (20) to each other. When the reaction chambers (20) are referred to as N-th chambers (N is a natural number) in the order adjacent to the sample inlet (11), the sample movement path (40) connects the N-th chamber and the N+1-th chamber. Here, “in the order adjacent to the sample inlet (11)” means the order in which the sample passes through the reaction chambers (20) sequentially through the sample movement path (40) from the reaction chamber (20) in which the sample first arrives after being injected through the sample inlet (11). According to the present embodiment, the sample movement path (40) includes a first flow path (41), a second flow path (42), and a first connecting flow path (43).

[0070] The first flow path (41) has one end connected to the Nth chamber and the other end extends toward the N+1th chamber. The second flow path (42) has the other end connected to the N+1th chamber and the other end extends toward the Nth chamber. The first connecting flow path (43) connects the first and second flow paths (41, 42) to each other. That is, one end of the first connecting flow path (43) is connected to the other end of the first flow path (41), and the other end of the first connecting flow path (43) is connected to one end of the second flow path (42). According to the present embodiment, the first and second flow paths (41, 42) include a bending portion (44) that changes the flow direction of the sample. At least one bending portion (44) may be provided.

[0071] According to the present embodiment, as illustrated in FIG. 3, during the process of moving from the first flow path (41) to the first connecting flow path (43) and during the process of moving from the first connecting flow path (43) to the second flow path (42), the sample is designed to change direction three times. However, the number of bending portions (44) is not limited thereto and may be formed in various ways. The bending portions (44) prevent the sample from moving between adjacent reaction chambers (20) when the sample is injected into all reaction chambers (20) provided in the body portion (100) and reacts with the reagent, so that the reaction result of each reaction chamber (20) is reliably secured without being mixed with the reaction result of another reaction chamber (20).

[0072]

[0073] The chamber unit (300) for testing the treatment solution is provided in the body unit (100) and includes a test chamber (30) in which a reaction reagent is received for testing the treatment solution and the reaction reagent reacts with the treatment solution injected through the treatment solution inlet (12). The reaction reagent is provided to detect whether the treatment solution contains a specific target genetic material. For example, the reaction reagent may be manufactured in a form that includes a specific target genetic material to determine whether the treatment solution is contaminated with the specific target genetic material. The reaction reagent may be contained in and fixed to the test chamber (30).

[0074] In the present embodiment, the treatment solution may be a solution such as a buffer used to extract nucleic acids from a sample. The sample is pretreated so that nucleic acids can be separated from the collected specimen before performing PCR. The pretreatment includes injecting a predetermined lysis buffer, such as a lysis buffer, into the biological sample to destroy the cell wall and leak the nucleic acids. The lysis buffer is a lysis buffer used to destroy the cell wall. The sample with the exposed nucleic acids is introduced into the reaction chamber. According to the present embodiment, the treatment solution inspection chamber unit can check for contamination of the treatment solution used for pretreatment of the sample, etc. Although the present embodiment has described the process of extracting nucleic acids using a lysis buffer as an example, it is of course also possible to check for contamination of the treatment solution used in the process of extracting nucleic acids by a nucleic acid extraction method using magnetic beads, a column-based nucleic acid extraction method, an automated extraction method, etc.

[0075] According to an embodiment of the present invention, the treatment solution may include not only those used for sample pretreatment, but also all treatment solutions used for sample treatment. For example, it may include sample dilution buffers such as DW and TE buffer, master mixes, etc. Meanwhile, the master mix may be fixed to the reaction chamber (20) and the inspection chamber (30) or may be mixed and added externally.

[0076] As illustrated in FIG. 2, the inspection chamber (30) includes an inlet portion (31) into which a treatment liquid is introduced, a well portion (32) formed by recessing from the lower surface of the body portion (100) toward the upper surface so that the treatment liquid and a reaction reagent react, and an outlet portion (33) through which the treatment liquid flows out from the well portion (32). The reaction reagent that reacts with the treatment liquid can be accommodated and fixed in the well portion (32). According to the present embodiment, the inspection chamber (30) is formed substantially similarly to the reaction chamber (20).

[0077] The chamber unit (300) for testing the treatment solution may include at least one testing chamber (30). When the chamber unit (300) for testing the treatment solution includes a plurality of testing chambers (30), the chamber unit may include a path for introducing the treatment solution and a testing chamber (30) including a reaction reagent for detecting whether the treatment solution includes a specific target genetic material. The embodiment according to FIG. 2 has four testing chambers (30), and each testing chamber (30) may be connected by a treatment solution movement path (50). Of course, according to another embodiment, such as FIG. 6, only one testing chamber (30) may be provided.

[0078] Referring to FIG. 2, a plurality of inspection chambers (30) are provided in the body portion (100). The inspection chambers (30) are connected to each other by a treatment liquid flow path (50) so that the treatment liquid flows in one direction. That is, the treatment liquid flow path (50) and the inspection chamber (30) are connected in series. The treatment liquid injection port (12) for injecting the treatment liquid is connected to both ends of the inspection chambers (30) connected in series. A user can inject the treatment liquid to be inspected using any one of the treatment liquid injection ports (12).

[0079] According to the present embodiment, when a plurality of inspection chambers (30) are provided, the treatment liquid movement path (50) for connecting the inspection chambers (20) to each other includes at least one curved portion in the section connecting the inspection chambers (30) and the adjacent inspection chambers (30). It is preferable that the treatment liquid movement path (50) be formed to be less than 0.55 mm to prevent excessive pressure from being generated when the treatment liquid moves, thereby inducing smooth movement. The inspection chamber (30) is provided in the form of a groove that is long in one direction.

[0080] The inlet portion (31) of the inspection chamber (30) includes an expansion portion (311) whose width gradually widens as it approaches the inspection chamber (30). The expansion portion (311) is formed to be tapered, so that when the treatment liquid flows into the inspection chamber (30), it is slowed down by the widening section and fills the internal space of the inspection chamber (30). The outlet portion (33) of the inspection chamber (30) includes a narrowing portion (331) whose width narrows as it moves away from the inspection chamber (30) as the treatment liquid flows out of the inspection chamber (30). The flow speed of the treatment liquid is increased by the narrowing portion (331), so that the treatment liquid can easily move to another adjacent inspection chamber (30).

[0081] The bottom surface of the above-mentioned inspection chamber (30) is formed deeper than the bottom surface of the above-mentioned treatment liquid movement path (50). The above-mentioned inlet (31) has an inclined surface (34) connected to the bottom surface of the above-mentioned inspection chamber (30), and the above-mentioned outlet (33) has an inclined surface (34) connected to the bottom surface of the above-mentioned inspection chamber (30) like the above-mentioned inlet (31). The bottom surface of the above-mentioned inspection chamber (30) is formed deeper than the bottom surface of the above-mentioned treatment liquid movement path (50) to form sufficient space for fixing the reaction reagent, and the reaction reagent is fixed to the deeper bottom to improve the reactivity with the treatment liquid. When a fluorescent substance is used, the deepened well portion (32) provides an effect that makes it easier to identify the fluorescent substance.

[0082] According to the present embodiment, the treatment liquid movement path (50) is provided to connect the plurality of inspection chambers (30) to each other. When the inspection chambers (30) are referred to as N-th chambers (N is a natural number) in the order adjacent to the treatment liquid injection port (12), the treatment liquid movement path (50) connects the N-th chamber and the N+1-th chamber. Here, “in the order adjacent to the treatment liquid injection port (12)” means the order in which the treatment liquid passes when it sequentially passes through the inspection chambers (30) through the treatment liquid movement path (50) from the inspection chamber (30) in which it first arrives after being injected through the treatment liquid injection port (12). According to the present embodiment, the treatment liquid movement path (50) includes a third path (51), a fourth path (52), and a second connecting path (53).

[0083] The third flow path (51) has one end connected to the Nth chamber and the other end extends toward the N+1th chamber. The second flow path (52) has the other end connected to the N+1th chamber and one end extends toward the Nth chamber. The second connecting flow path (53) connects the third and fourth flow paths (51, 52) to each other. That is, one end of the second connecting flow path (53) is connected to the other end of the third flow path (51), and the other end of the second connecting flow path (53) is connected to one end of the fourth flow path (52). According to the present embodiment, the third and fourth flow paths (51, 52) include a bending portion (54) that changes the flow direction of the treatment liquid. At least one bending portion (44) may be provided.

[0084] According to the present embodiment, as illustrated in FIG. 3, the treatment liquid is designed to change direction three times during the process of moving from the third flow path (51) to the second connecting flow path (53) and during the process of moving from the second connecting flow path (53) to the fourth flow path (52). However, the number of banding portions (54) provided in the treatment liquid movement path (50) is not limited thereto and may be formed in various ways. The banding portions (54) prevent the sample from moving between adjacent test chambers (30) when the treatment liquid is fully injected into the test chambers (30) provided in the body part (100) and reacts with the reagent, so that the reaction results of each test chamber (30) can be reliably secured without being mixed with the reaction results of other test chambers (30). The test chamber (30) may contain a plurality of reaction reagents for detecting a specific target genetic material.

[0085]

[0086] According to this embodiment, a cover part (60) is coupled to the lower surface of the body part (100).

[0087] The cover part (60) is provided to close the sample inspection chamber part (200) and the treatment liquid inspection chamber part (300), which are provided in a groove shape from the lower surface toward the upper surface of the body part (100). When the cover part (60) is attached to the lower surface of the body part (100) and the body part (100) is positioned so that the cover part (60) faces downward, the cover part (50) forms the lower surface of the sample inspection chamber part (200) and the treatment liquid inspection chamber part (300). That is, when the sample and treatment liquid injected through the sample inlet (11) and the treatment liquid inlet (12) are transferred to the sample inspection chamber part (200) and the treatment liquid inspection chamber part (300), the lower surfaces of the sample movement path (40) and the treatment liquid movement path (50) are formed by the cover part (50).

[0088] According to the present embodiment, the cover part (60) is provided in the form of a film so as to be disposed on the lower side of the body part (100) and to receive heat from a heating device for performing PCR, and is attached to the lower surface of the body part (100). The cover part (60) is made of a thin film so as to easily receive heat from the heating device, and may be made of a metal with excellent heat transfer efficiency or a polymer material including nano carbon. Meanwhile, the sample and the treatment liquid can be automatically moved in the sample movement path (40) and the treatment liquid movement path (50) without using any special power. An adhesive area (R) is provided on the lower surface of the body part (100) so that the film-shaped cover part (60) can be attached. The adhesive area (R) is formed to be wider than the width of the sample movement path (40) and the treatment liquid movement path (50) so as to prevent leakage when the treatment liquid or sample is injected. The above adhesive area (R) is formed at the edge of the reaction chamber (20), the inspection chamber (30), the sample movement path (40), the treatment liquid movement path (50), and the edge of the body (100), and may have a width of 0.8 mm to 1.0 mm. If the width of the adhesive area (R) is less than 0.8 mm, the adhesive strength of the cover part (50) is reduced, and if it exceeds 1.0 mm, the spatial arrangement of the reaction chamber (20) is restricted.

[0089] Meanwhile, the multifunctional PCR test chip (1001, 1002, 1003) according to the embodiment of the present invention is inserted into a PCR device to perform PCR, and the PCR device may include a main body in which the chip (1001, 1002, 1003) according to the present invention is mounted, a heating means provided inside the main body to provide heat to the chip (1001, 1002, 1003), a light source that irradiates light to the chip (1001, 1002, 1003), a camera that captures an image by photographing fluorescence when a reaction reagent and a sample react, and an analysis unit that analyzes the image of the camera. The present invention relates to the chip (1001, 1002, 1003), and a detailed description of the PCR device itself is omitted.

[0090]

[0091] (1) Regarding the first embodiment

[0092]

[0093] According to one embodiment of the present invention, the reaction chamber (20) and the inspection chamber (30) may be provided in multiple numbers. For example, the reaction chambers (20) may be aligned adjacent to each other, and the inspection chamber (30) may be provided at least one on each outermost side of the plurality of reaction chambers (20) aligned adjacent to each other. For example, referring to the multifunctional PCR inspection chip (1001) illustrated in FIG. 4, the sample inspection chamber unit (200) may include six reaction chambers (20), and the treatment solution inspection chamber unit (300) may include four inspection chambers (30). According to this embodiment, one of the inspection chambers (30) is adjacent to one side of the six reaction chambers (20) that are arranged in parallel, and the remaining three inspection chambers (30) are arranged in parallel adjacent to the last reaction chamber (20). At this time, three inspection chambers (30) can be aligned adjacent to each other.

[0094]

[0095] Specifically, the chip according to the embodiment of FIG. 4 can be utilized as follows. In the first inspection chamber (30), reaction reagents capable of detecting six target genes (hereinafter referred to as target 1 gene to target 6 gene) are inserted together, and in each of the second to seventh reaction chambers (20), reaction reagents capable of detecting one target gene are inserted.

[0096] That is, a reaction reagent for detecting the target 1 gene can be fixed in the second reaction chamber (20), a reaction reagent for detecting the target 2 gene can be fixed in the third reaction chamber (20), a reaction reagent for detecting the target 3 gene can be fixed in the fourth reaction chamber (20), a reaction reagent for detecting the target 4 gene can be fixed in the fifth reaction chamber (20), a reaction reagent for detecting the target 5 gene can be fixed in the sixth reaction chamber (20), and a reaction reagent for detecting the target 6 gene can be fixed in the seventh reaction chamber (20). In addition, target 1, 2 genes and a reaction reagent for detecting them can be fixed in the eighth test chamber (30), target 3, 4 genes and a reaction reagent for detecting them can be fixed in the ninth test chamber (30), and target 5, 6 genes and a reaction reagent for detecting them can be fixed in the tenth test chamber (30).

[0097] Of course, the position of the reaction reagent for detecting the target gene can be changed depending on the situation, and the number of the reaction chamber (20) and the test chamber (30) can also be changed depending on the design. In this state, the treatment solution (dissolution buffer) is injected into the first test chamber (30), filling 8 to 10 test chambers (30), and the sample is injected into the second reaction chamber (20), filling up to the seventh reaction chamber (20), and then PCR is performed.

[0098] Since the treatment solution that is not contaminated with the Target 1 to Target 6 genes will not show any reaction in the first test chamber (30), it can be verified that the treatment solution is not contaminated (NC). In addition, if the chip inserted into the PCR device undergoes amplification and amplification proceeds well in the 8th to 10th test chambers (30), it can be verified that the PCR device or reaction reagents are functioning properly.

[0099] Meanwhile, an IC (internal control) may be injected into the reaction chamber (20) to verify whether nucleic acid extraction has been performed properly. The IC is intended to verify the unique genes that must be included in the sample, and the validity of nucleic acid extraction from the sample can be determined by determining whether the nucleic acid extracted by the IC satisfies quantitative standards. The IC may be fixed to one of the reaction chambers (20) or may be directly injected into the reaction chamber (20) by the user prior to performing PCR.

[0100] Meanwhile, two types of reaction reagents can be fixed to each reaction chamber to detect two target genetic materials. According to the embodiment of Fig. 4, since six reaction chambers (20) are provided, a total of 12 target genes can be detected. Accordingly, in order to detect whether the treatment solution is contaminated with the 12 target genes, reaction reagents capable of reacting to 1 to 12 target genetic materials can be fixed to the first test chamber (30) to serve as NC (negative control), and test chambers 8 to 10 (30) can serve as PC (positive control) for verifying PCR equipment and reagents.

[0101] In this way, the validity of the PCR equipment, reaction reagents, and / or PCR reaction can be verified through the inspection chamber (30) and the reaction chamber (20). In this state, when the treatment solution or DW is injected into the inspection chamber (30), if there is no reaction to the target gene contained in the inspection chamber (30), the treatment solution can be judged as not contaminated (NC: negative control), and if the PCR amplification is performed smoothly, the reagent can be judged as functioning properly (PC: positive control).

[0102] In addition, a sample can be introduced into the reaction chamber (30) to test whether a target gene is detected. That is, when a treatment solution is introduced into the test chamber (30) and a sample is introduced into the reaction chamber (30) to be tested, the presence of contamination in the treatment solution, the smooth operation of the reagent, and the test for the sample can be performed on a single chip.

[0103]

[0104] (2) Regarding the second embodiment

[0105]

[0106] FIG. 6 illustrates a multifunctional PCR test chip (1002) according to another embodiment of the present invention. According to this embodiment, as illustrated in FIG. 6, the configuration of the upper surface of the body portion (100) is different from the embodiment of FIG. 4 in that the positions of the sample inlet (11) and the treatment solution inlet (12) are changed. As illustrated in FIG. 7, the multifunctional PCR test chip (1002) according to this 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 treatment solution test chamber portion (300) are provided on the lower surface of the body portion (100), wherein the sample test chamber portion (200) includes nine reaction chambers (20), and the treatment solution test chamber portion (300) includes one test chamber (30).

[0107] For each reaction chamber (20), two types of reaction reagents can be fixed to detect two target genetic materials. That is, in this case, a total of 18 target genes can be detected. Accordingly, in order to detect whether the treatment solution is contaminated with the 18 target genes, reaction reagents capable of reacting to the 18 target genetic materials can be fixed to one of the test chambers (30). Of course, a reaction reagent for detecting one target genetic material can also be fixed to each reaction chamber (20).

[0108] In this state, when the treatment solution or DW is injected into the test chamber (30) and there is no reaction to the 18 target genes, the treatment solution is judged to be uncontaminated, enabling NC (negative control), and the sample can be injected into the reaction chamber (20) to test whether the target gene is detected. In other words, when the sample is injected into the reaction chamber (20) and tested, the presence or absence of contamination of the treatment solution can be performed on a single chip.

[0109] Additionally, an IC (internal control) may be injected into one of the reaction chambers (20) to verify whether nucleic acid extraction has been performed properly. The IC is a primer or probe that can detect a unique gene possessed by a living organism, and can confirm whether nucleic acid extraction has been performed properly from the sample. By using the IC, it is possible to determine whether the extracted nucleic acid satisfies a quantitative standard, thereby determining the validity of nucleic acid extraction from the sample. The IC may be fixed to one of the reaction chambers (20) or may be directly injected into the reaction chamber (20) by the user before performing PCR.

[0110] Meanwhile, in the present embodiment, when chambers 1 and 10 are connected to each other to form a chamber unit (200) for examining a treatment solution, and chambers 2 and 9 are connected to each other to form a chamber unit (200) for examining a sample, chamber 10 can be utilized to function as a PC (posotive control) as in the first embodiment.

[0111]

[0112] (3) Regarding the third embodiment

[0113]

[0114] According to a multifunctional PCR test chip (1003) according to another embodiment of the present invention, the sample test chamber parts (200) may be provided in multiple numbers, and the treatment liquid test chamber parts (300) may also be provided in multiple numbers. When performing PCR, any one of the multiple sample test chamber parts (201, 202) and any one of the multiple treatment liquid test chamber parts (301, 301) may be combined with each other to form one set. That is, according to the present embodiment, a plurality of sets in which the sample test chamber parts (201, 202) and the treatment liquid test chamber parts (301, 302) are combined may be provided.

[0115] Referring to FIG. 9, the sample inspection chamber unit (200) includes a first sample inspection chamber unit (201) in which a plurality of reaction chambers (200) are provided adjacent to each other, and a second sample inspection chamber unit (202) in which a plurality of reaction chambers (20) are provided adjacent to each other and separated from the first sample inspection chamber unit (201). In addition, the treatment solution inspection chamber unit (300) may include at least one first inspection chamber (301) adjacent to the first sample inspection chamber unit (201) and at least one second inspection chamber (302) adjacent to the second sample inspection chamber unit (202). According to the present embodiment, the first sample inspection chamber unit (201) and the second sample inspection chamber unit (202) are arranged adjacent to each other, but are arranged between the first inspection chamber (301) and the second inspection chamber (302).

[0116] According to the present embodiment, as illustrated in FIG. 9, the first inspection chamber (301) and the first sample inspection chamber unit (201) constitute one set, and the second sample inspection chamber unit (202) and the second inspection chamber (302) constitute another set.

[0117] Specifically, the first sample inspection chamber unit (201) has four reaction chambers (20) arranged adjacently and connected to each other by a sample movement path (40). The second sample inspection chamber unit (202) is not connected to the first sample inspection chamber unit (202) and is configured to include four reaction chambers (20) adjacent to each other. The second sample inspection chamber unit (202) is also connected by a sample movement path (40).

[0118] Of course, the number of reaction chambers (20) constituting the first and second sample inspection chamber parts (201, 202) is not limited to the four described above and can be varied. Referring to FIG. 9, the first inspection chamber (301) is arranged adjacent to the outermost left side of the first sample inspection chamber part (201), and the second inspection chamber (302) is arranged adjacent to the outermost right side of the second sample inspection chamber (202). The first inspection chamber (301) and the first sample inspection chamber part (201) and the second inspection chamber (302) and the second sample inspection chamber part (202) are formed symmetrically to each other. Of course, the first inspection chamber (301) may be provided on the outermost right side of the first sample inspection chamber section (201), or the second inspection chamber (302) may be provided on the outermost left side of the second sample inspection chamber section (202).

[0119] The first and second inspection chambers (301, 302) are each provided with two treatment solution inlets (12). The first sample inspection chamber unit (201) and the second sample inspection chamber unit (202) are each provided with two sample inlets (11) at both ends. According to the present embodiment, the first inspection chamber (301) is used to inspect whether the treatment solution or DW for the treatment of the first sample is contaminated, and the first sample inspection chamber unit (201) can be used to inspect whether the target gene is detected when the first sample is introduced into the reaction chamber (20). In addition, the second inspection chamber (302) is used to inspect whether the treatment solution or DW for the treatment of the second sample is contaminated, and the second sample inspection chamber unit (202) can be used to inspect whether the target gene is detected when the second sample is introduced into the reaction chamber (20).

[0120] Referring back to FIG. 9, two types of reaction reagents can be fixed to each of the reaction chambers (20) numbered 2 to 5 constituting the first sample inspection chamber unit (201) for detecting two target genetic materials. That is, in this case, a total of eight target genes can be detected. Accordingly, in order to detect whether the treatment solution for processing the first sample is contaminated with the eight target genes, reaction reagents capable of reacting to the eight target genetic materials can be fixed to the first inspection chamber (301). Of course, a reaction reagent for detecting one target genetic material may also be fixed to each reaction chamber (20) of the first sample inspection chamber unit (201).

[0121] In this state, if the treatment solution or DW for processing the first sample is injected into the first test chamber (301) and there is no reaction to the eight target genes, the treatment solution is judged to be uncontaminated, enabling NC (negative control), and the first sample is injected into the first sample test chamber (201) to test whether the target gene is detected.

[0122] That is, when the first sample is introduced into the reaction chamber (20) and tested, it is possible to perform the contamination of the treatment solution that processes the first sample on a single chip. In addition, an IC (internal control) can be injected into any one of the reaction chambers (20) to verify whether nucleic acid extraction has been performed properly. As described above, the validity of nucleic acid extraction of the sample can be determined by determining whether the extracted nucleic acid satisfies the quantitative standard by the IC. The IC can be fixed to any one of the reaction chambers (20) that constitute the first sample inspection chamber unit (201), or can be directly introduced into the reaction chamber (20) by the user before performing PCR.

[0123] Meanwhile, in the present embodiment, chambers 1 and 5 may be connected to each other to form a first inspection chamber (301), and chambers 2 to 4 may be connected to each other to form a first sample inspection chamber section (201). In this case, chamber 5 may be utilized to function as a PC (posotive control) as in the first embodiment.

[0124] The second sample testing chamber unit (202), similar to the first sample testing chamber unit (201), can have two types of reaction reagents fixed to each of the 6th to 9th reaction chambers (20) for detecting two target genetic materials. That is, in this case, a total of eight target genes can be detected. The eight target genes may be the same targets as those of the first sample testing chamber unit (201). In order to detect whether the treatment solution for processing the second sample is contaminated with the eight target genes, reaction reagents capable of reacting to the eight target genetic materials can be fixed to the second testing chamber (302). Of course, a reaction reagent for detecting one target genetic material may be fixed to each reaction chamber (20) of the second sample testing chamber unit (20).

[0125] In this state, when the treatment solution or DW for processing the second sample is injected into the second test chamber (302), if there is no reaction to the eight target genes, the treatment solution is judged to be uncontaminated, enabling NC (negative control), and the second sample is injected into the second sample test chamber (202) to test whether the target gene is detected. That is, when the second sample is injected into the reaction chamber (20) and tested, the contamination of the treatment solution for processing the second sample can be performed on a single chip.

[0126] Additionally, an IC (internal control) may be injected into one of the reaction chambers (20) to verify whether nucleic acid extraction has been successfully performed. As described above, the IC can be used to determine whether the extracted nucleic acid satisfies quantitative criteria, thereby determining the effectiveness of nucleic acid extraction from the sample. The IC may be fixed to one of the reaction chambers (20) constituting the second sample testing chamber (201), or may be directly injected into the reaction chamber (20) by the user prior to performing PCR.

[0127] Meanwhile, in the present embodiment, chambers 6 and 10 may be connected to each other to form a second inspection chamber (302), and chambers 7 to 9 may be connected to each other to form a second sample inspection chamber unit (202). In this case, chamber 6 may be utilized to function as a PC (posotive control) as in the first embodiment.

[0128] In this way, according to the present embodiment, a chip capable of simultaneously performing tests on different samples is provided. According to the present embodiment, two samples can be tested with a single chip, and at this time, the presence or absence of contamination in each processing solution used to process the two samples can be simultaneously checked. Of course, depending on the target genetic material to be detected, the number of the reaction chambers and the test chambers, or the type and number of reaction reagents fixed to the reaction chambers and the test chambers can be varied.

[0129]

[0130] By this process, the present invention provides the effect of enabling multiplex testing by detecting multiple types of target genes with a single sample injection, and simultaneously checking for contamination of a processing solution, such as a buffer, used to extract nucleic acids from a sample, on a single chip. That is, a chamber unit for testing a sample, including a plurality of reaction chambers into which a sample is injected, and a chamber unit for testing a processing solution, which tests for contamination by injecting a processing solution, etc. to enable NC (negative control), are formed separately, so that testing of the sample and the processing solution can be performed simultaneously.

[0131] In addition, the chamber for testing the treatment solution is configured with multiple test chambers to enable confirmation of whether the reaction reagent is functioning properly (PC: positive control), thereby providing the effect of simultaneously performing a test using a sample, a test for contamination of the treatment solution, and a check of whether the reaction reagent is functioning properly.

[0132] In addition, the present invention was developed for the purpose of simultaneous multiplex diagnosis with high sensitivity and specificity. In order to solve the problem that in the existing multiplex PCR diagnosis, a large number of primer probes that detect several targets are gathered in the same space, which leads to a decrease in sensitivity and specificity due to interference and competitive reactions and different reaction characteristics among them, the detection zones of each target are spatially separated, so that interference and competitive reactions among them can be eliminated. In addition, when the reaction reagent is contained and fixed in the reaction chamber and reacts with the sample, the sensitivity can be improved when confirming the test result.

[0133]

[0134] Above, the present invention has been described in detail with reference to preferred embodiments, but the present invention is not limited to the above embodiments, and many modifications can be provided within a range that does not depart from the scope of the present invention.

Claims

1. A body part made of a light-transmitting material and having a sample inlet for injecting a sample and a treatment solution inlet for injecting a treatment solution for processing the sample; A chamber for sample inspection provided in the above body portion, which includes a reaction chamber in which a reaction reagent for detecting a target genetic material is received and in which a sample introduced through the sample introduction port reacts with the reaction reagent; and A multifunctional PCR test chip characterized by including a chamber for testing a treatment solution provided in the body portion and including a test chamber in which the reaction reagent is received and the treatment solution injected through the treatment solution injection port reacts with the reaction reagent.

2. In paragraph 1, The above reaction chamber, An inlet into which the sample is introduced; A reaction well formed by the sample reacting with the reaction reagent and sinking from the lower surface of the body toward the upper surface; and A multifunctional PCR test chip characterized in that it includes an outlet through which the sample flows out from the reaction well.

3. In paragraph 2, A multifunctional PCR test chip characterized in that the inlet and outlet of the reaction chamber are formed at an angle.

4. In paragraph 2, The above reaction chamber is provided in multiple numbers in the above body part, A multifunctional PCR test chip characterized in that the reaction chambers are connected to each other by a sample movement path so that the sample flows in one direction between the reaction chambers.

5. In paragraph 2, The above inlet portion includes an expansion portion whose width widens in the direction in which the sample enters the reaction well portion, A multifunctional PCR test chip characterized in that the above-mentioned discharge portion includes a reduced portion whose width narrows in the direction in which the sample flows into the reaction well portion.

6. In paragraph 4, The sample transfer path connects the Nth chamber and the N+1th chamber when the reaction chamber is called the Nth chamber (N is a natural number) in the order adjacent to the sample inlet. The above sample transfer path is: The first end is connected to the Nth chamber; A second end connected to the N+1 chamber; and A first connecting passage connecting the first euro and the second euro; A multifunctional PCR test chip, characterized in that the first and second euros include at least one banding portion so that the flow direction of the sample is changed.

7. In paragraph 1, On the lower surface of the above body part, a chamber part for examining the sample, which is provided in a groove shape from the lower surface toward the upper surface, and a cover part for closing the chamber part for examining the treatment liquid are combined, A multifunctional PCR test chip characterized in that the cover part forms the bottom surface of the sample inspection chamber part and the treatment solution inspection chamber part while the body part is positioned so that the cover part faces downward.

8. In paragraph 7, A multifunctional PCR test chip characterized in that the cover part is provided in the form of a film and is attached to the lower surface of the body part so as to be positioned on the lower side of the body part and transmit heat from a heating device for performing PCR.

9. In paragraph 1, A multifunctional PCR test chip characterized in that the above test chamber is provided in multiple numbers.

10. In paragraph 9, A multifunctional PCR test chip characterized in that the test chambers are connected to each other by a treatment liquid flow path so that the treatment liquid flows in one direction.

11. In paragraph 10, The above treatment liquid movement path connects the M chamber and the M+1 chamber when the inspection chamber is called the M chamber (M is a natural number) in the order adjacent to the treatment liquid injection port. The above treatment liquid movement path is: A third Euro, which is connected to the M chamber at one end; A fourth end connected to the M+1 chamber; and A second connecting passage connecting the third and fourth euros; A multifunctional PCR test chip, characterized in that the third and fourth euros include at least one banding portion so that the flow direction of the sample is changed.

12. In paragraph 4, The above plurality of reaction chambers are aligned adjacent to each other, A multifunctional PCR test chip characterized in that a plurality of the above test chambers are provided, and at least one test chamber is provided on the outermost sides of the plurality of reaction chambers that are aligned adjacent to each other.

13. In paragraph 1, The sample inspection chamber section includes a first sample inspection chamber section in which a plurality of reaction chambers are provided adjacent to each other, and a second sample inspection chamber section in which a plurality of other reaction chambers are provided adjacent to each other, separated from the first sample inspection chamber section. A multifunctional PCR test chip, characterized in that the chamber section for testing the treatment solution includes at least one first test chamber adjacent to the chamber section for testing the first sample and at least one second test chamber adjacent to the chamber section for testing the second sample.

14. In paragraph 13, A multifunctional PCR test chip characterized in that the first sample test chamber and the second sample test chamber are arranged adjacent to each other, but are arranged between the first test chamber and the second test chamber.

15. In paragraph 1, A multifunctional PCR test chip characterized in that reaction reagents for detecting at least two different target genes are fixed in the reaction chamber and the test chamber, thereby enabling multiple diagnosis for each sample.

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