PCR Plate, Nucleic Acid Extraction Cartridge, and Polymerase Chain Reaction Device Containing the Same

Abstract

The PCR plate according to the present invention includes: a body portion having one or more reaction wells; an insertion portion extending from the body portion, inserted into a nucleic acid extraction cartridge, and having an injection port into which a nucleic acid solution is injected; a flow path portion allowing the nucleic acid solution to flow from the injection port to the reaction wells; and a blocking portion attached to the body portion for blocking backflow of the nucleic acid solution from the reaction wells to the flow path portion side.

Description

Technical Field

[0001] The present invention relates to a PCR plate, a nucleic acid extraction cartridge, and a polymerase chain reaction device including the same, and more particularly, to a PCR plate, a nucleic acid extraction cartridge, and a polymerase chain reaction device including the same that can extract, amplify, and detect the amplified product of nucleic acid in real time.

Background Art

[0002] POC (Point of care) diagnostic technology for accurately and rapidly diagnosing a patient's disease regardless of time and location has attracted attention as a very important technology for evidence-based precision medicine. By examining all infectious pathogens that cause disease symptoms based on symptoms such as cough, diarrhea, high fever, and genital abnormalities within a short time at once, the causative pathogen is confirmed, and symptom-based on-site diagnosis for prescribing the optimal antibiotic and therapeutic agent is the core new technology of future precision medicine and is developing while many studies are being conducted. Such on-site diagnostic technology has the advantage of being quickly and accurately diagnosed by non-experts on-site, like existing pregnancy test kits for confirming pregnancy and blood glucose meters for confirming blood glucose. Currently, multiplex testing methods that can simultaneously test various pathogens have been developed, and molecular diagnostic technology has attracted attention as a core technology of future medicine that can utilize these technologies to accurately identify the cause of infectious diseases, enable optimal prescribing, treat diseases early, significantly shorten the patient's recovery period, improve the quality of medical care, and reduce medical costs.

[0003] However, the current molecular diagnostic system takes more than 3 hours to confirm the results and must be used by trained experts. Therefore, for POC molecular diagnosis required on-site, it is essential to develop an automated small device that can perform complex nucleic acid extraction processes and real-time gene amplification tests automatically and must be easily operable by non-experts.

[0004] As a typical molecular diagnostic method, there is a method using polymerase chain reaction (PCR, Polymerase Chain Reaction, hereinafter referred to as "PCR"). Since the invention of polymerase chain reaction (PCR) by Kary Mullis in 1985, specific DNA can be amplified quickly and easily, so it has been widely used in molecular biology and molecular diagnosis. When using PCR / RT-PCR, it is possible to confirm whether specific DNA / RNA exists in a biological sample. Therefore, this technology is often used in the diagnosis of pathogenic microorganism infections such as viruses. While this PCR / RT-PCR technology has evolved into real-time quantitative PCR (Realtime PCR, Quantitative PCR), it can confirm the results simultaneously with the end of PCR. This not only simplifies the testing process and significantly reduces the testing time but also accurately quantifies the number of pathogens. Therefore, it is used as a standard diagnostic method for monitoring the treatment effects of viruses such as HIV, HCV, and HBV. In addition, since the PCR / RT-PCR technology can detect gene expression patterns and gene mutations related to specific diseases, it is used as the most important technology for disease diagnosis.

[0005] To perform such PCR, a nucleic acid extraction step is required to remove each substance that inhibits the PCR reaction from the biological sample and extract pure nucleic acids. The nucleic acid extraction process consists of multiple steps and requires skilled techniques for biological samples and nucleic acid extraction operations. When performed manually, problems such as contamination due to operator errors may occur. Therefore, molecular diagnosis is mostly carried out using automated nucleic acid extraction equipment.

[0006] To detect the PCR reaction and reaction products, a real-time quantitative PCR device must be equipped. Therefore, in the past, molecular diagnosis was mainly performed in large hospitals and specialized clinical testing institutions.

[0007] Through recent research and development, various automated systems that automate the entire process of nucleic acid extraction, PCR reaction, and reaction product detection and each device using this have been developed, enabling easy use of PCR without specialized techniques.

[0008] However, each of the existing devices has problems such as being overly expensive, requiring a long processing time, and being difficult to perform various inspections at once.

[0009] To explain the basic principle of PCR, after separating the DNA double helix into single strands by heating it to 95°C, the reaction solution is cooled to the annealing temperature, and the complementary primers in the PCR reaction solution are selectively hybridized to both ends of the site to be amplified. Then, DNA polymerase sequentially links the four types of nucleotide triphosphates, A, G, T, and C, complementary to each single strand, and repeats the reaction to form a double helix. PCR is an experimental reaction that repeats the heating and cooling of the PCR reaction solution 30 to 45 cycles (n) to geometrically amplify only 2 n copies of a specific DNA double helix. The RT-PCR reaction is extended to a method for detecting RNA by synthesizing cDNA through a reverse transcription reaction and then becoming amplifiable through PCR.

[0010] To explain the principle of newly developed real-time quantitative PCR for the full-scale application of PCR in molecular diagnosis, for the quantitative analysis of DNA amplified using the PCR reaction, a substance that generates fluorescence in proportion to the amount of DNA is added to the PCR reaction solution. Then, the fluorescence is measured for each cycle, the cycle at which the critical fluorescence value is detected is found, and from this, the concentration of the initial target nucleic acid is quantitatively measured.

[0011] Since the invention of PCR, while various application technologies have been developed, through the genome project, the base sequences of many pathogens and disease-related genes have been known. Molecular diagnosis that amplifies such disease-related DNA / RNA base sequences and diagnoses them qualitatively and quantitatively has developed rapidly. Existing PCR takes about 2 hours in the temperature cycle, so various methods that can perform PCR more quickly and accurately for on-site diagnosis have been continuously developed (Lab Chip, 2016, 16, 3866 - 3884).

[0012] In order to perform a PCR reaction within a short time, the temperature of the reaction solution must be changed rapidly. Also, in order to amplify only the desired target in an accurate PCR reaction, each primer must be designed to specifically attach to the desired target, and the annealing temperature must be accurately adjusted in the PCR temperature cycling reaction.

[0013] For this purpose, a micro PCR reaction vessel with as small a heat capacity as possible and good heat transfer has been developed, compared to the 0.2 ml and 0.5 ml reactors commonly used in existing laboratories. Each of these microreactors uses less reaction solution and has a large surface area, so heat is rapidly transferred and fast heating and cooling are possible. 10 μl of PCR solution was placed in a thin reaction groove of 40 μm to 80 μm formed on a silicon wafer with a size of 17×15 mm and covered with a glass plate to maintain a large surface area (>100 mm 2 / 10 μl), but it was not possible to shorten the time to about 3 minutes per cycle using an existing Peltier type thermal block (Clin.Chem.40 / 9,1815 - 1818(1994)).

[0014] In an initial PCR reaction apparatus, in order to rapidly thermally cycle the PCR reactor, a method of repeatedly immersing and transferring the PCR reactor in a high-temperature water bath and a low-temperature water bath was developed (Turbo Thermalcycler. Bioneer Corp. Daejeon). Such a PCR apparatus that circulates the reactor in regions with different temperatures is a spatial movement method, and by immersing the reactor in a thermostatic water bath where the temperature is accurately maintained in advance, it has the advantage of being able to perform the PCR reaction rapidly and accurately. However, since a large number of thermostatic water baths are required, the equipment is large and difficult to maintain, and PCR equipment that adopts a time-difference temperature cycling method of changing the temperature over time using a Peltier element or the like with a fixed block has become the mainstay.

[0015] PCR methods using microfluidic channels have also been developed for spatial temperature cycling and time-lag temperature cycling. The spatial temperature cycling method can be broadly divided into an open type that continuously flows out in a FIFO (First-In-First-Out) manner and a closed type that repeatedly moves through different temperature zones. The open type was developed in 1994 by Nakano et al. by winding a capillary around a cylindrical block having compartments with different temperatures and continuously flowing a PCR solution through it (Biosci. Biotech. Biochem., 58(2), 349 - 352, 1994). In 1998, Kopp et al. confirmed that PCR can be advanced by passing a 10 μl solution through a microfluidic channel PCR device that repeatedly passes through high and low temperature zones in the form of a microfluidic channel, with a cycle of 4.5 seconds for 20 cycles (Science 280 1046 - 1048, 1998).

[0016] The background art of the present invention is disclosed in Korean Patent Publication No. 10 - 2016 - 0067872 (published on June 14, 2016, title of the invention: Analytical unit, analytical apparatus, operating method of the analytical unit, and manufacturing method of the analytical unit for performing polymerase chain reaction).

[0017] In the conventional Korean Registered Patent No. 10 - 2105558, a high-speed polymerase chain reaction analysis plate is disclosed. However, when using the disclosed analysis plate, there may be a problem that amplification products between each reaction well are mixed during the polymerase chain reaction process, reducing the accuracy of the experiment. To solve such problems, an improved PCR plate that can quickly perform quantitative and qualitative analysis of a large number of targets is required.

Summary of the Invention

Problems to be Solved by the Invention

[0018] The present invention has been made to solve the above problems, and an object of the present invention is to perform automatic target nucleic acid detection through extraction of nucleic acids from a biological sample, PCR reaction, and excitation light in various wavelength bands and corresponding fluorescence scanning, to be able to inspect a large number of targets by a single operation, and to be easy to use. In particular, it is to provide a PCR plate, a nucleic acid extraction cartridge, and a polymerase chain reaction device including the same that can obtain accurate results within a short time.

[0019] Another object of the present invention is to enable rapid and accurate PCR by being able to rapidly and repeatedly apply the temperature required for the heat denaturation process and the accurate temperature required for the binding process to the reaction object in the temperature adjustment process required for the PCR process, and to provide a PCR plate, a nucleic acid extraction cartridge, and a polymerase chain reaction device including the same that can maximize the reliability of the reaction.

Means for Solving the Problems

[0020] The PCR plate according to the present invention includes a body portion having one or more reaction wells; an insertion portion extending from the body portion, inserted into a nucleic acid extraction cartridge, and having an injection port into which a nucleic acid solution is injected; a flow path portion that allows the nucleic acid solution to flow from the injection port to the reaction wells; and a blocking portion attached to the body portion that blocks backflow of the nucleic acid solution from the reaction wells to the flow path portion side.

[0021] In the present invention, the body portion may include a body frame portion having one or more of the reaction wells; a blocking accommodation portion formed in a concave shape in the body frame portion for accommodating the blocking portion; a connecting flow path portion connecting the blocking accommodation portion and the reaction wells; a storage portion formed in a concave shape in the body portion, communicating with the flow path portion, and receiving the supply of the nucleic acid solution; and a flow path guide portion communicating with the storage portion and guiding the nucleic acid solution to flow toward the blocking portion side.

[0022] In the present invention, the connecting flow path portion may include a bottleneck portion formed with a narrow width from the blocking portion toward the reaction well side.

[0023] In the present invention, the blocking accommodation part may be divided into a plurality of parts by a partition wall part formed along the length direction of the storage part, and the blocking part may be attached to each of the divided blocking accommodation parts.

[0024] In the present invention, the blocking part may be configured to include an elastically deformable material.

[0025] In the present invention, the body part may include: a body frame part having one or more of the reaction wells; a blocking accommodation part formed in a concave shape in the body frame part, circularly formed so as to accommodate the circular blocking part corresponding to each of the reaction wells; and a connecting flow path part connecting the blocking accommodation part and the reaction well.

[0026] In the present invention, the blocking part may be configured to include an elastically deformable material.

[0027] In the present invention, a sheet part may be heat-sealed to the reaction well.

[0028] The nucleic acid extraction cartridge according to the present invention may include: a cartridge cover having accommodation parts with a large number of partition structures containing solutions necessary for DNA extraction; and a cartridge main body part coupled to the lid part by an insertion structure and provided with a reaction accommodation part for reacting or purifying a solution drawn from the accommodation part with a sample.

[0029] In the present invention, the cartridge cover may be in contact with one side surface of the accommodation part of the cartridge main body part and may include a rubber part containing an elastically deformable material.

[0030] In the present invention, a liquid leakage prevention rubber containing an elastic material and preventing leakage of the nucleic acid solution may be attached to the nucleic acid extraction cartridge.

[0031] The nucleic acid extraction cartridge assembly according to the present invention includes a nucleic acid extraction cartridge; and a PCR plate that receives a nucleic acid solution from the nucleic acid extraction cartridge and accommodates the solution in a reaction well containing a PCR mixture dry matter. The nucleic acid extraction cartridge includes a cartridge cover having a housing portion with a number of partition structures containing solutions necessary for DNA extraction; and a cartridge main body portion coupled to the lid portion by an insertion structure and provided with a reaction housing portion for reacting or purifying a solution drawn from the housing portion with a sample. The PCR plate includes a body portion having one or more of the reaction wells; an insertion portion extending from the body portion, inserted into the nucleic acid extraction cartridge, and having an injection port into which the nucleic acid solution is injected; a flow path portion that enables the nucleic acid solution to flow from the injection port to the reaction well; and a blocking portion attached to the body portion for blocking backflow of the nucleic acid solution from the reaction well to the flow path portion side.

[0032] In the present invention, the cartridge cover may include a rubber portion in contact with one side surface of the housing portion of the cartridge main body portion and containing an elastically deformable material.

[0033] In the present invention, a liquid leakage prevention rubber containing an elastic material and preventing leakage of the nucleic acid solution may be attached to the nucleic acid extraction cartridge.

[0034] The polymerase chain reaction device according to the present invention includes a nucleic acid extraction cartridge; and a PCR plate that receives a nucleic acid solution from the nucleic acid extraction cartridge and accommodates the nucleic acid solution in a reaction well containing a dried PCR mixture. The nucleic acid extraction cartridge includes a cartridge cover having a housing portion with a number of partition structures containing solutions necessary for DNA extraction; and a cartridge main body portion that is coupled to the lid portion by an insertion structure and is provided with a reaction housing portion for reacting or purifying a solution drawn from the housing portion with a sample. The PCR plate includes a body portion having one or more of the reaction wells; an insertion portion extending from the body portion, inserted into the nucleic acid extraction cartridge, and having an injection port into which the nucleic acid solution is injected; a flow path portion that allows the nucleic acid solution to flow from the injection port to the reaction well; and a blocking portion that is attached to the body portion and blocks backflow of the nucleic acid solution from the reaction well to the flow path portion side.

[0035] In the present invention, the cartridge cover can include a rubber portion that contacts one side surface of the housing portion of the cartridge main body portion and includes an elastically deformable material.

[0036] In the present invention, a liquid leakage prevention rubber containing an elastic material may be attached to the nucleic acid extraction cartridge to prevent leakage of the nucleic acid solution.

Advantages of the Invention

[0037] According to the present invention, extraction of nucleic acids from a biological sample, PCR reaction, and real-time reaction product detection through excitation light in various wavelength bands and corresponding fluorescence scanning can be automated, various tests can be performed by a single operation, the use is simple, and accurate results can be obtained in a short time.

[0038] In addition, in the PCR plate, backflow of the dried PCR mixture from the reaction well can be prevented, so that contamination of adjacent reaction wells by mixing of the dried PCR mixture can be prevented.

[0039] In addition, according to the present invention, in the temperature control required for the PCR process, the reliability of the reaction can be maximized by enabling the accurate temperature required for the heat denaturation stage and binding to be rapidly applied to the reaction target in real time at once, thereby enabling accurate PCR.

[0040] That is, in the case of the conventional temperature control method of increasing the temperature while moving the reaction solution, the temperature cannot be increased uniformly, which is disadvantageous for the PCR reaction. In the method of sequentially increasing the temperature while the reaction solution moves, temperature uniformity cannot be achieved simultaneously for the entire reaction product. Therefore, the present invention eliminates the problem of an increased substrate for other reactions occurring, maintains the temperature range set in the heating block in a constant temperature state, and increases the temperature by directly pressurizing the entire reaction solution, thereby very efficiently implementing the temperature increase required for the PCR reaction.

[0041] Furthermore, in order to minimize the time delay in the process of changing the temperature from high to low, block-shaped heating block structures are arranged side by side and separated. When pressurizing the PCR plate, the position of the heating block is changed so that pressurization is performed in real time by heating blocks having individually different temperatures, and the problem caused by the time delay required in the temperature change process can be epoch-makingly solved.

[0042] Furthermore, the PCR plate is embodied in an insertion type structure into the nucleic acid extraction cartridge. The nucleic acid extraction cartridge is commonly used, and while storing the PCR plates used in various test kits in a narrow space, the appropriate PCR plate can be inserted and used as needed during the test. One reaction well provided in the PCR plate can analyze up to six fluorescence values, and if necessary, the number of reaction wells of the PCR plate can be increased up to eight, so that all pathogenic bacteria that may be contained in the biological sample of the patient related to the symptoms can be amplified and detected, thereby enabling a multiplex molecular diagnostic test based on the symptoms.

[0043] Also, according to the present invention, when the constant temperature plate is divided into regions having a gradient of a first temperature and a second temperature, and the heating block is pressurized through the drive module, it moves so that a region having a set temperature (the first temperature or the second temperature) corresponding to the temperature of the heating block corresponds, and contact and pressurization are simultaneously performed on the upper and lower surfaces of the PCR plate. As a result, it is possible to realize twice the efficiency compared to the constant temperature plate method maintained at a single temperature.

[0044] In addition, in implementing the movable constant temperature plate structure, by using a sliding tape in a configuration that performs a driving operation, the reliability of the product configuration and movement can be ensured, and since a method of simultaneously heating the plate contained in the target on the upper and lower surfaces is adopted, the inspection time can be shortened.

Brief Description of the Drawings

[0045]

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Embodiments for Carrying Out the Invention

[0046] Hereinafter, with reference to the accompanying drawings, an embodiment of a PCR plate, a nucleic acid extraction cartridge, and a polymerase chain reaction apparatus including the same according to the present invention will be described. In such a process, the thickness of each line and the size of each component shown in the drawings may be exaggerated for clarity and convenience of explanation.

[0047] Also, each term described below is a term defined in consideration of the functions in the present invention, and this may vary depending on the intentions or conventions of users and operators. Therefore, these terms should be defined based on the content throughout this specification.

[0048] FIG. 1 is a block diagram showing the main component configuration of a polymerase chain reaction apparatus according to an embodiment of the present invention. FIGS. 2 to 7 are diagrams for explaining the structure of a temperature control module in an embodiment of the present invention. FIGS. 8 to 13 are diagrams showing an embodiment of a PCR plate applied to an embodiment of the present invention. FIGS. 14 to 19 are diagrams showing another embodiment of a PCR plate applied to an embodiment of the present invention. FIGS. 20 to 23 are conceptual diagrams for explaining the structure and operation of a thermostatic plate and a horizontal movement drive module applied to an embodiment of the present invention. FIG. 24 is a perspective conceptual diagram of a nucleic acid extraction cartridge according to an embodiment of the present invention, showing a structure in which the above-described PCR plate is inserted and coupled. FIG. 25 is a separated perspective view of FIG. 24. FIG. 26 is a diagram showing the internal structure of a cartridge cover in the structure of FIG. 24. FIG. 27 is a perspective view showing the coupled state of the structure of FIG. 24. FIG. 28 is an assembled perspective view of another embodiment of a nucleic acid extraction cartridge according to an embodiment of the present invention. FIG. 29 is a cross-sectional view of another embodiment of a nucleic acid extraction cartridge according to an embodiment of the present invention. FIGS. 30 to 32 are diagrams showing the lower operating state of a cartridge structure according to an embodiment of the present invention. FIG. 33 is a diagram showing the overall structure and arrangement configuration of an apparatus constituting a polymerase chain reaction apparatus according to an embodiment of the present invention described above. FIG. 34 is an enlarged view of a combined arrangement diagram of a main part of an embodiment of the present invention in FIG. 33. FIG. 35 is a vertical cross-sectional conceptual diagram of a part of FIG. 34, presenting the arrangement of the main part configuration. FIG. 36 is a side perspective cross-sectional view of FIG. 35. FIG. 37 is a perspective conceptual diagram of a nucleic acid extraction cartridge according to another embodiment of the present invention. FIG. 38 is a cross-sectional view of FIG. 37. FIG. 39 is a diagram showing punching a liquid leakage prevention rubber with a punch in FIG. 38. FIG. 40 is a diagram showing pressurizing a liquid leakage prevention rubber with a pressing jig in FIG. 39. FIG. 41 is a diagram showing that the liquid leakage prevention rubber prevents residual liquid leakage. FIG. 42 is a diagram showing still another embodiment of a PCR plate. FIG. 43 is a cross-sectional view of FIG. 42. FIG. 44 is a diagram showing that a sheet is attached to the PCR plate. FIG. 45 is a diagram showing a test for confirming the presence or absence of mixing of a solution between reaction wells during PCR. FIG. 46 is a diagram showing a mixing flow test of reaction wells in a PCR plate.

[0049] Referring to FIG. 1, the polymerase chain reaction apparatus according to an embodiment of the present invention enables accurate PCR by applying the temperature required for the heat denaturation process and the exact temperature required for the annealing process to the reaction object in real time without a time difference during the temperature control process necessary for the PCR process, and can maximize the reliability of the reaction. A temperature control module embodied by a heating block structure that contacts the PCR reaction plate and applies a specific temperature can be provided.

[0050] The temperature control module according to the present invention minimizes the time delay required to embody the temperature from the first temperature to the second temperature, which is relatively low, or vice versa from the second temperature to the first temperature, which is relatively high. By changing the positions of the heating blocks respectively set to the first temperature and the second temperature, pressurization to the PCR reaction plate is performed in real time, and problems due to the time delay required in the temperature change process can be epoch-makingly solved.

[0051] Furthermore, in the present invention, it can be embodied to further include a constant temperature plate structure that is disposed below the PCR plate and operates in a structure that horizontally moves in a sliding manner. In this case, by the constant temperature plate structure that maintains the temperature of the PCR plate at the first temperature or the second temperature, the required time required for applying the temperature change conditions can be minimized, and the reaction rate can be maximized.

[0052] The polymerase chain reaction device according to an embodiment of the present invention includes a PCR plate 200 that receives a nucleic acid solution from a nucleic acid extraction cartridge 100 and accommodates it in a reaction well W containing a PCR mixture dry product, and is disposed on one side of the PCR plate 200 and adjacent to the reaction well W. A temperature control module 300 including a pair of heating blocks 310 and 320 capable of applying different temperatures to each other and capable of horizontal movement and vertical movement. The PCR plate 200 includes a body portion 210 having one or more of the reaction wells W, an insertion portion 220 extending from the body portion 210, inserted into the nucleic acid extraction cartridge 100, and having an injection port h1 into which a nucleic acid solution is injected, and from the injection port h1 to the reaction well W. It may include a flow path portion 230 that allows the nucleic acid solution to flow, and a blocking portion 240 that is attached to the body portion 210 and blocks the backflow of the nucleic acid solution from the reaction well W to the flow path portion 230 side.

[0053] With the above-described configuration, the present invention provides the convenience of allowing a desired sample to be introduced through the nucleic acid extraction cartridge 100 even by a non-expert and freely performing nucleic acid extraction, and at the same time, in the temperature cycle required for amplification applied to the PCR plate 200. Quick and precise temperature control is possible using the heating block 310, 320 structures capable of directly applying the target temperature to the reaction solution in the PCR plate 200 through pressure bonding with a thin film.

[0054] In addition, a polymerase chain reaction (hereinafter referred to as "PCR") device embodied in one system can be provided so that a scanning module can perform a detection operation on a reaction product through excitation light in various wavelength bands and corresponding fluorescence scanning at the lower part of the PCR plate 200 in real time.

[0055] FIGS. 2 to 7 are diagrams for explaining the structure of the temperature control module 300 in the present invention.

[0056] FIGS. 2 and 3 are schematic perspective views of the temperature control module of the present invention.

[0057] Referring to FIGS. 2 and 3, the temperature control module 300 extracts the nucleic acid of the biological sample, receives and stores the PCR (polymerase chain reaction) premix or nucleic acid solution from the nucleic acid extraction cartridge 100 while mixing with the polymerase, and performs a function of performing constant temperature control on the PCR plate 200.

[0058] The temperature control module 300 may be provided with a first pressing surface G1 corresponding to the surface of the reaction well W embodied in the PCR plate 200, and a first heating block 310 maintained at a temperature set within the range of the temperature required for heat denaturation (hereinafter, "first temperature") by a heating unit. In addition, a second heating block 320 may be provided at a position spaced apart from the position corresponding to the first heating block 310, with a second pressing surface G2 corresponding to the surface of the reaction well W, and maintained at a temperature set within the range of the temperature required for annealing (hereinafter, "second temperature") by a heating unit. In particular, the structures of the first heating block 310 and the second heating block 320 may be embodied so as to enable horizontal movement and vertical movement.

[0059] In one embodiment, the first heating block 310 and the second heating block 320 have a three-dimensional structure and may have a flat pressing surface on the lower surface. The first heating block 310 and the second heating block 320 may be arranged spaced apart from each other and may have temperatures in different temperature ranges.

[0060] As shown in FIGS. 2 and 3, the first heating block 310 and the second heating block 320 are arranged opposite to each other in a facing structure, and the upper surface as a whole is embodied in a structure for performing a flat pressing function, and the upper part thereof may be provided with a rectangular parallelepiped three-dimensional structure. The embodiment of the rectangular parallelepiped three-dimensional structure is one example, and it can be said that any three-dimensional shape as long as it has a flat pressing surface for pressing is included in the gist of the present invention.

[0061] Further, the first heating block 310 and the second heating block 320 may be arranged on the side surfaces, and the adjacent surfaces may be configured to be spaced apart from each other, and each may be maintained to have a different set temperature.

[0062] That is, the first temperature of the first heating block 310 may be set in the range of 94° C. to 96° C. corresponding to the temperature applied to the thermal denaturation step of separating double-stranded DNA (including DNA extracted from a biological sample), and in an example of the present invention, it may be maintained at 95° C.

[0063] Furthermore, the second temperature of the second heating block 320 may be the temperature required for the primer binding step (annealing) to allow the primer to bind to the separated template DNA, and may be set in the range of 50° C. to 65° C., and in an example of the present invention, it may be maintained at 55° C.

[0064] The first heating block 310 and the second heating block 320 are not configured to accommodate water or a heat transfer fluid inside, but are embodied in a structure having a metal body with a large heat capacity and good heat transfer efficiency, and can be constantly maintained at the set temperature by an internal heating unit. For this purpose, a temperature sensor is installed inside, and the heating unit must be controlled through temperature adjustment so that a constant temperature can be maintained.

[0065] That is, when a PCR premix or a nucleic acid solution is injected into the PCR plate 200, at the time when the application of the first temperature is required, the first heating block 310 comes adjacent to the surface of the PCR plate 200 while moving horizontally. That is, since the first pressing surface G has a flat plate structure, it is possible to heat the entire surface of the PCR plate 200 at the same temperature and the same pressing force at the same time, and uniform temperature transfer to the entire sample becomes possible.

[0066] Also, when it is necessary to apply the second temperature required for the bonding stage, the second heating block 320 is positioned above the PCR plate 200 while horizontally moving, enabling heating of the entire surface of the PCR plate 200 simultaneously at the same temperature and the same pressing force.

[0067] That is, since there is no need for a separate time to prepare for the set temperature reaction and it can be driven in a manner that enables heating the entire surface of the PCR plate 200 simultaneously at the same temperature and the same pressing force by a simple horizontal movement, a rapid and precise PCR reaction can be derived compared to the existing method for controlling the set temperature.

[0068] In addition, since the first heating block 310 and the second heating block 320 are set to different temperatures, a cooling fan unit 340 capable of embodying a cooling effect in the separation space between the two structures is provided, taking into account that the second heating block 320 can be constantly heated by the radiant heat and conductive heat of the first heating block 310.

[0069] Since it is important for the second heating block 320 to be able to be maintained at a relatively second temperature, for example, an annealing temperature of 55°C, it can be provided with a heat dissipation type cooling pattern at the upper part that minimizes the heat interference of the first heating block 310 and allows excessive heat to be easily dissipated by the cooling fan unit. As such an example, in the present invention, the second heating block 320 can further include a temperature adjustment pattern unit 321 embodied on the side surface of the second pressing surface G2. The temperature adjustment pattern unit 321 has a structure in which a large number of protruding patterns are embodied on the upper part, and since it can increase the heat dissipation efficiency while expanding the contact surface area with air, it is advantageous for maintaining a constant low temperature.

[0070] Unlike the method of moving the reaction sample that embodies the PCR reaction or moving it to another heating region through time setting, the heating block structure is applied so that the reaction sample can be fixed and the temperature can be uniformly increased simultaneously overall from above, enabling the accurate transmission of the first temperature and the second temperature.

[0071] In addition, the first heating block 310 and the second heating block 320 in the present invention can be interlocked with a drive module 330 that embodies a horizontal movement or a vertical movement operation. The drive module 330 includes guide members 331 and 332 that penetrate the first heating block 310 and the second heating block 320, and the first heating block 310 and the second heating block 320 can move up and down along the guide members 331 and 332.

[0072] That is, the first heating block 310 and the second heating block 320 in the present invention are arranged separately from each other and are configured to move up and down while intersecting each other by the operation of the drive module 330. Furthermore, elastic members S1 and S2 arranged below the guide members 331 and 332 are further included, and when the heating blocks 310 and 320 press the PCR plate 200, a buffering action can be performed by applying an appropriate elastic force (see Fig. 5).

[0073] Also, the polymerase chain reaction apparatus according to an embodiment of the present invention can further include a thermostatic plate 350 that is interlocked with the temperature control module 300.

[0074] As shown in FIGS. 2 and 3, the constant temperature plate 350 is disposed at the lower part of the heating blocks 310 and 320 that constitute the temperature control module 300. After the PCR plate 200 enters, when the first heating block 310 and the second heating block 320 of the temperature control module 300 pressurize the PCR plate 200 through horizontal movement and vertical movement, it can function to have the same temperature as that of the first heating block 310 and the second heating block 320.

[0075] For this purpose, the constant temperature plate 350 can be further configured to include a horizontal movement drive module 400 that moves horizontally under the PCR plate 200.

[0076] For this purpose, the constant temperature plate 350 can be further configured to include a horizontal movement drive module 400 that moves horizontally under the PCR plate 200.

[0077] As shown in FIGS. 2 and 3, the horizontal movement drive module 400 can include a moving bar 420 coupled to one end of the constant temperature plate 350, a drive motor unit 410, and a conversion plate 430 that converts the rotational force of the drive motor unit 410 into the horizontal movement force of the moving bar 420.

[0078] Such a horizontal movement drive module 400 enables the constant temperature plate 350 to move horizontally in the lower direction of the temperature control module 300 described above. In particular, the constant temperature plate 350 according to an embodiment of the present invention can be implemented in a structure partitioned into a first region heated to a first temperature and a second region heated to a second temperature separated from the first region.

[0079] In particular, in an embodiment of the present invention, the temperature control module 300 and the constant temperature plate 350 can be implemented as an integrated unit through the guide members 331 and 332.

[0080] That is, when the horizontal movement drive module 400 is driven, the temperature control module 300 including the thermostatic plate 350 and the heating blocks 310 and 320 can be moved together.

[0081] In this case, the thermostatic plate 350 includes a first region heated to a first temperature and a second region heated to a second temperature separated from the first region. The first pressing surface G1 of the first heating block 310 is arranged to correspond to the upper part of the first region. On the upper and lower surfaces of the PCR plate 200, the PCR plate 200 is arranged between the thermostatic plate 350 and the heating blocks 310 and 320, and pressure can be applied at the same temperature simultaneously.

[0082] That is, in the embodiment of the present invention, when the thermostatic plate 350 is divided into regions having a gradient of a first temperature and a second temperature, and the heating blocks are pressed through the horizontal movement drive module 400, the regions having set temperatures (the first temperature or the second temperature) corresponding to the temperatures of the heating blocks 310 and 320 move horizontally in a sliding structure so as to correspond, and by simultaneously contacting and pressing the upper and lower surfaces of the PCR plate 200, the efficiency is twice that of the thermostatic plate 350 method maintained at a single temperature.

[0083] FIG. 4 is a cross-sectional view of the temperature control module in FIG. 3 seen from the rear, and FIG. 5 is a cross-sectional view of the temperature control module seen from the front.

[0084] Thus, the temperature control module 300 of the present invention is provided with a drive module 330 that realizes the horizontal movement or vertical movement of the first heating block 310 and the second heating block 320, enabling the operation of such a heating module to be automated.

[0085] Referring to FIGS. 2 to 5, the drive module 330 moves the first heating block 310 and the second heating block 320 up and down. At the same time, the horizontal movement drive module 400 horizontally moves the first heating block 310 and the second heating block 320 so that the portion in contact with the surface of the reaction well W on the PCR plate 200 can be changed to the first pressing surface G1 or the second pressing surface G2.

[0086] The first heating block 310 and the second heating block 320 are arranged side by side in a separated state. Each of them is provided with guide grooves 312 and 322 (see FIG. 2) in a through structure. The first heating block 310 and the second heating block 320 are placed on guide members 331 and 332 that penetrate the guide grooves 312 and 322. Through this, the first heating block 310 and the second heating block 320 move up and down along the guide members 331 and 332, and the PCR plate 200 can be pressed from above.

[0087] Of course, in this case, a thermostatic plate 350 is arranged below the first heating block 310 and the second heating block 320, corresponding to the regions having the same temperature as the first temperature or the second temperature provided in each heating block, so that the PCR plate can be pressed from above and below.

[0088] As described above, the temperature control module 300 in the present invention realizes the advantage that the first temperature and the second temperature can be directly applied to the entire surface of the PCR plate for the entire PCR target in the PCR plate 200, and excellent effects can be realized in terms of the application speed side and the reaction efficiency side.

[0089] In addition, the structure of the heating block of the temperature control module 300 is positioned at the upper part where horizontal movement is always possible, and it has a structure that descends to the lower part only when it is pressure-bonded to the PCR plate 200.

[0090] To implement such a structure, it can include a first elastic member S1 that is arranged in a structure inserted inside the drive frame and has a restoring force that always raises upward when not crimped. Also, in the present invention, when applying pressure in contact with the PCR plate 200, in order to prevent applying an excessive pressing force, a second elastic member 335 that transmits the pressing force is provided (Fig. 5). In the illustrated embodiment, the second elastic member 335 is implemented as a leaf spring structure, and when the first and second heating blocks 310 and 320 are pressed in the lower direction, it exerts a certain buffering force so that an excessive pressing force is not applied to the surface of the PCR plate 200 and can be controlled accordingly.

[0091] Also, in the present invention, the PCR plate 200 has a form of a plate-shaped structure with reaction wells implemented on the upper surface. In this case, the PCR plate 200 can be further configured to include a constant temperature plate 350 structure at the lower part so that it can be maintained within a certain temperature range, for example, the second temperature (e.g., 55°C).

[0092] This is because when raising the temperature while closely attaching the first heating block at the first temperature (e.g., 95°C) or closely attaching the second heating block at the second temperature (e.g., 55°C) by the temperature control module 300 of the present invention, the internal amplification efficiency becomes much better only when the PCR plate 200 quickly reaches the target temperature.

[0093] Therefore, in an embodiment of the present invention, it can be further configured to include a constant temperature plate 350 that is arranged below the PCR plate 200 and maintains the temperature of the PCR plate 200 at the second temperature.

[0094] In particular, providing the constant temperature plate 350 structure may be implemented in a way of fixedly mounting and applying a constantly set temperature. However, as described above, the constant temperature plate 350 itself can be divided into regions for applying the first temperature and the second temperature while partitioning it, and can be implemented in a structure where the constant temperature plate can be horizontally moved.

[0095] FIG. 6 shows a state in which in the structure of FIG. 2, the thermostatic plate 350 is horizontally moved through the horizontal movement drive module 400 and enters below the temperature control module. FIG. 7 shows a state in which the thermostatic plate 350 is horizontally moved outward in the operation of FIG. 6 to change the temperature region.

[0096] That is, in the structure of FIG. 6, when the first heating block 310 is arranged to apply the first temperature, the first region in the thermostatic plate 350 horizontally moves and is arranged below the PCR plate together with the first heating block, and the first heating block 310 corresponds to face the upper surface of the PCR plate 200. Then, as shown in FIG. 7, when the second region horizontally moves and is arranged below the PCR plate together with the second heating block, the second heating block 320 horizontally moves and operates corresponding to face the upper surface of the PCR plate 200, which is shown.

[0097] The horizontal movement operation of the thermostatic plate 350 is implemented in a sliding manner and can be implemented to move while contacting a sliding tape that contacts the side surface of the thermostatic plate 350.

[0098] Also, referring to FIGS. 6 and 7, this is a conceptual diagram showing the bottom surface of the temperature control module 300 according to the present invention. As shown in the figure, the thermostatic plate 350 is arranged between the PCR plate 200 and the lower scanning module (not shown: reference numeral 500 in FIG. 1), and excitation light irradiated from the scanning module is transmitted to the PCR plate 200, and a plurality of light transmission portions H through which the light of the scanning module 500 is guided so that fluorescence can be detected may be provided in a through structure.

[0099] Therefore, in the present invention, in the integrated system equipped with the above-described scanner, nucleic acid extraction, PCR process, and detection process can be implemented in one system, and a separated PCR plate structure can be implemented so as to be applicable to various disease diagnoses.

[0100] Figures 8 to 13 are diagrams showing an embodiment of the PCR plate 200 applied to the present invention.

[0101] Referring to FIG. 8 and referring to the conceptual diagrams of FIGS. 2 and 3 described above, the PCR plate 200 according to the present invention includes a body portion 210, an insertion portion 220, a flow path portion 230, and a blocking portion 240.

[0102] The body portion 210 includes one or more reaction wells W (W1... Wn). The reaction well W receives a PCR (polymerase chain reaction) premix or a nucleic acid solution (hereinafter referred to as "nucleic acid solution") from the nucleic acid extraction cartridge 100 on the plate-shaped surface and accommodates a primer, a primer / probe, or a dried PCR mixture containing a primer / probe.

[0103] The insertion portion 220 extends from the body portion 210, is inserted into the nucleic acid extraction cartridge 100, and includes an injection port h1 into which the nucleic acid solution is injected. The insertion portion 220 extends from one end of the body portion 210 and is coupled in a structure that is inserted into the nucleic acid extraction cartridge 100.

[0104] In particular, an insertion portion 220 having an injection port h1 into which the nucleic acid solution is injected so as to be drawn from the nucleic acid extraction cartridge 100 can be implemented. Further, the PCR plate 200 can be implemented in a structure that is connected to a flow path portion 230 provided on the surface of the body portion 210 so as to be connected to the injection port h1 and connected to a number of reaction wells W.

[0105] In the present invention, in the case of the reaction well W, in the structure shown in FIG. 8, it is implemented in a structure having eight reaction wells W1 to W8, but it is not limited thereto, and it is natural that it can also be implemented in a structure having one or more numbers, and the structure of the reaction well W can also be implemented in a concave pattern structure while processing the surface of the body portion 210.

[0106] In particular, in the embodiments of the present invention, as shown in FIG. 8, a structure is provided to partition a region of a certain reaction well W in the surface region of the body portion 210, so that the primer provided in a dried state in the reaction well W and the nucleic acid solution injected from the nucleic acid extraction cartridge 100 can be dispersed and mixed in the reaction well W.

[0107] The flow path portion 230 enables the nucleic acid solution to flow from the injection port h1 to the reaction well W. The blocking portion 240 is attached to the body portion 210 and blocks the backflow of the nucleic acid solution from the reaction well W to the flow path portion 230 side. The blocking portion 240 is configured to include an elastically deformable material. The elastically deformable material of the blocking portion 240 may be made of rubber, silicone, or the like.

[0108] The body portion 210 includes a body frame portion 211, a blocking housing portion 213, a connecting flow path portion 215, a storage portion 217, and a flow path guide portion 219. The body frame portion 211 includes one or more reaction wells W (W1... Wn).

[0109] The blocking housing portion 213 is formed in a concave shape in the body frame portion 211 and houses the blocking portion 240. The blocking housing portion 213 is formed in a concave shape corresponding to the size of the blocking portion 240 so as to be able to house the blocking portion 240.

[0110] The connecting flow path portion 215 connects the blocking housing portion 213 and each reaction well W. The nucleic acid solution can be supplied to the reaction well W through the connecting flow path portion 215.

[0111] The storage portion 217 is formed in a concave shape in the body portion 210, communicates with the flow path portion 230, and receives the supply of the nucleic acid solution. The storage portion 217 receives and stores the supply of the nucleic acid solution so as to be able to supply it to each reaction well W.

[0112] The flow path guide portion 219 communicates with the storage portion 217 and guides the nucleic acid solution to flow toward the blocking portion 240 side.

[0113] The connection channel portion 215 includes a bottleneck portion 215a that is formed with a narrow width from the blocking portion 240 toward the reaction well W side. The bottleneck portion 215a is formed such that the width suddenly narrows in the connection channel portion 215, and the portion where the blocking portion 240 is accommodated in the blocking accommodation portion 213 protrudes slightly.

[0114] Referring to FIG. 10, the blocking portion 240 is inserted into the blocking accommodation portion 213 while being pressurized from the upper side to the lower side, and the width of the bottleneck portion 215a of the connection channel portion 215 is formed to be narrow.

[0115] Referring to FIG. 11, when a transparent film is sealed and adhered to the bottleneck portion 215a by pressurization, the four sides are pressurized in the connection channel portion 215 by the blocking portion 240, and the flow of the nucleic acid solution is blocked. Therefore, the flow of the nucleic acid solution is blocked by the weak pressure difference between adjacent reaction wells W.

[0116] Referring to FIG. 12, when the nucleic acid solution is continuously injected into the blocking portion 240, the blocking portion 240 is pushed while passing through the flow path guide portion 219 from the storage portion 217, and the nucleic acid solution flows into each reaction well W.

[0117] Referring to FIG. 13, after the nucleic acid solution flows into each reaction well W, when the pressure increases due to the temperature change of each reaction well W, the blocking portion 240 is pressurized in the reaction well W, and the blocking portion 240 is brought into close contact with and pressurizes the flow path guide portion 219, so that the nucleic acid solution can be blocked from flowing backward from the reaction well W to the flow path portion 230 side.

[0118] Further, the PCR plate 200 is configured to include a cover member (not shown) that seals the upper portions of a large number of reaction wells W, and a film material made of a transparent material having light transmissivity can be applied to the cover member.

[0119] When the cover member embodies the inside of the reaction well W as a cavity while being in close contact with the surface of the reaction well W, the PCR (polymerase chain reaction) premix and the nucleic acid solution injected later from the nucleic acid extraction cartridge 100 are injected into the inside of the reaction well W while pushing out the air layer existing in the cavity.

[0120] In particular, in the present invention, as shown in the structure of FIG. 8, the flow path portion 230 connected to the reaction well W in the body portion 210 is embodied to extend from the injection port h1 through the body portion 210 to the end portion of the body portion 210, and at the end portion, it can be embodied to be respectively connected to the ends of a number of reaction holes in the opposite direction to the insertion portion 220.

[0121] That is, as shown in FIG. 2, when the PCR premix and the nucleic acid solution are injected into the nucleic acid extraction cartridge 100 through the injection port h1 provided at the lower part of the insertion portion, the flow path portion 230 is embodied in the direction x1 crossing the body portion 210, branched left and right from the end point of the body portion 210, and can be embodied to be connected to the inlets of the respective reaction wells W. The reason for forming the flow path in this way is that a minute air layer exists inside the region of the reaction well W sealed by the cover member, so that the injected PCR premix and nucleic acid solution rise from the upper region with respect to the center line Cx of the body portion as shown in the structure of FIG. 8, and the air layer rises to the lower region Cb of the body portion 210.

[0122] Therefore, the mixture for which the PCR reaction is carried out is arranged in the upper region Ca of the reaction well W relatively, and this is because the regions where the heating blocks 310 and 320 of the present invention perform pressurization and the region where the scanner module performs detection are in the upper region Ca as shown in FIG. 8 due to the characteristics of the equipment, and all of the detection precision, the efficiency of the PCR reaction, and the efficiency of temperature control can be enhanced.

[0123] In the present invention, the PCR plate 200 can be embodied with a synthetic resin material having high light transmittance. This is to enhance the detection efficiency by configuring it with a material having high light transmittance due to the function of the scanner module described above.

[0124] For such a material, various synthetic resin materials such as transparent PP, PE, PPA, PMMA, and PC can be applied, but it is not necessarily limited thereto, and it can be said that any material that can ensure a certain light transmittance is applicable.

[0125] However, the PCR plate 200 is adapted to maintain a constant temperature by a heat source applied from the lower temperature control plate 350, and for the efficiency of temperature maintenance of the temperature control module directly applied to the PCR reaction product including at least one of the PCR (polymerase chain reaction) premix or nucleic acid solution filled in the reaction well W and the dried primer and probe, the thickness of the body portion 210 can be embodied in the range of 1.0 mm to 3.0 mm. When the thickness of the body portion 210 is less than 1.0 mm, the high-temperature heat for setting the first temperature is easily transmitted to the lower part of the body portion 210, and heat interference with the temperature control plate 350 occurs, so temperature control is not easy. When the thickness of the body portion 210 exceeds 3.0 mm, temperature control for the substance accommodated in the reaction well W is easy, but temperature control of the lower temperature control plate 350 becomes difficult, and there is a disadvantage that it is difficult to maintain a constant temperature.

[0126] That is, in the present invention, as described with reference to FIGS. 4 and 5, the first pressing surface G1 or the second pressing surface G2 in contact with the surface of the reaction well W while horizontally moving the first heating block 310 and the second heating block 320 is pressed in a structure in contact with the upper surface of the partition pattern and the cover member covering the partition pattern, and the temperature of the reaction product is controlled while setting the temperature to the first temperature or the second temperature.

[0127] When performing temperature cycling on the PCR plate 200, in the temperature control module 300 with the structure shown in FIGS. 2 and 3, to raise the temperature to the first temperature, the first heating block 310 moves horizontally so as to face the upper surface of the PCR plate 200. On the lower surface of the PCR plate 200, after the first region of the constant temperature plate 350 moves horizontally together with the first heating block 310, the first heating block 310 moves downward while making contact and applying pressure. To lower the temperature to the second temperature, the upper surface of the PCR plate 200 moves horizontally so as to face the second heating block 320. On the lower surface of the PCR plate 200, after the second region of the constant temperature plate 350 moves horizontally together with the second heating block 320, the second heating block 320 moves downward while making contact and applying pressure, and it is driven so that heating and cooling are simultaneously performed on the upper and lower surfaces of the PCR plate 200. In the present invention, since the constant temperature plate 350 and the first and second heating blocks 310 and 320 are embodied in a structure that horizontally moves together, it is natural that the pressurization of the upper and lower parts of the PCR plate 200 can be realized at the same temperature simultaneously.

[0128] By such an operation, by simultaneously making contact and applying pressure to the upper and lower surfaces of the PCR plate 200, an efficiency twice that of the constant temperature plate 350 method maintained at a single temperature can be realized. Therefore, an advantage is realized in that the heating of the plate contained in the target is simultaneously realized on the upper and lower surfaces, and the inspection time can be reduced to 1 / 2.

[0129] FIGS. 14 to 19 are diagrams showing other embodiments of the PCR plate 200 applied to the present invention.

[0130] Referring to FIG. 14 and referring to the conceptual diagrams of FIGS. 2 and 3 described above, the PCR plate 200 of another embodiment according to the present invention includes a body portion 210, an insertion portion 220, a flow path portion 230, and a blocking portion 240. The descriptions of the body portion 210, the insertion portion 220, the flow path portion 230, and the blocking portion 240 of the PCR plate 200 are as described above.

[0131] In another embodiment of the PCR plate 200 of the present invention, the blocking accommodation portion 213 is divided into a plurality of parts by a partition portion 214 formed along the length direction of the storage portion 217. The blocking portion 240 is mounted on each of the blocking accommodation portions 213 divided by the partition portion 214.

[0132] Referring to FIG. 16, a plurality of blocking portions 240 are inserted and mounted on their respective blocking accommodation portions 213 from the upper side to the lower side. The elastically deformable blocking portion 240 is press-fitted into the blocking accommodation portion 213. The elastically deformable blocking portion 240 seals the blocking accommodation portion 213.

[0133] Referring to FIG. 17, after being sealed by the blocking portion 240, the flow due to the pressure difference in each blocking accommodation portion 213 is blocked by the partition portion 214.

[0134] Referring to FIG. 18, when the nucleic acid solution is injected, it pushes the blocking portion 240 while passing through the connecting flow path portion 215 via the storage portion 217, and the nucleic acid solution flows into each reaction well W.

[0135] Referring to FIG. 19, after the inflow of the nucleic acid solution, when the pressure due to the temperature change in each reaction well W increases, the blocking portion 240 is pressurized in the reaction well W. The blocking portion 240 can block the backflow of the nucleic acid solution while closely adhering to and pressurizing the connecting flow path portion 215.

[0136] FIGS. 20 to 23 are diagrams for explaining in detail the structure and operation method of such a thermostatic plate and the horizontal movement driving module.

[0137] FIG. 20 is a diagram showing the structure in which the thermostatic plate 350 is placed in FIGS. 2 and 3, and FIG. 22 is a diagram showing the structure in which only the thermostatic plate structure is separated.

[0138] Referring to FIGS. 20 and 21, the constant temperature plate 350 according to an embodiment of the present invention is disposed below the heating blocks 310 and 320 that constitute the temperature control module 300 shown in FIGS. 2 and 3. After the PCR plate 200 is disposed, when the first heating block 310 or the second heating block 320 of the temperature control module 300 presses the PCR plate 200 by horizontal movement and vertical movement, it can function to have the same temperature as the temperature of the first heating block 310 or the second heating block 320.

[0139] As shown in FIG. 20, the constant temperature plate 350 is provided with a partition Ss that partitions the first region a1 that maintains the first temperature and the second region a2 that maintains the second temperature. The first region a1 and the second region a2 are embodied in a structure that is connected based on both side ends a3 and a4 of the partition Ss.

[0140] Connectors Ca and Cb are mounted on one end of the constant temperature plate 350 so that power can be applied or a control signal can be transmitted.

[0141] In particular, the horizontal movement operation of the constant temperature plate 350 is embodied in a sliding manner, and it is embodied to move in contact with a sliding tape that contacts the side surface of the constant temperature plate 350. This not only simplifies the structure but also enhances the mobility.

[0142] In this case, the second region a2 portion may be provided with a light transmission portion H so that the detection light of the scanning module 500 that scans the concentration of the amplified reactant can pass through.

[0143] The first region a1 and the second region a2 are provided with temperature sensors Sa and Sb, and the temperature of the corresponding region can be measured and controlled.

[0144] FIG. 23 is a view showing the lower surface of FIG. 22, in which connection connectors Cc and Cd for applying a control signal and a power supply are provided, and temperature sensors Sa and Sb are provided so as to realize maintaining a constant temperature of a first temperature and a second temperature.

[0145] In order to maintain the temperature of the first region and the second region of the constant temperature plate 350, various heating means, such as a heating wire and a heating low antibody, can be used to mount various means inside the plate. However, in a preferred embodiment of the present invention, a circuit for an electrode and a temperature sensor is embodied in an epoxy printed circuit, and after applying a heat generating paint between each electrode, a metal plate corresponding to the first region and the second region is adhered so as to be in close contact with each temperature sensor and the heat generating paint, so as to realize such an effect.

[0146] In the present invention, the constant temperature plate 350 can be further configured to include a horizontal movement drive module 400 that moves horizontally below the PCR plate 200.

[0147] As shown in FIGS. 2 and 3, the horizontal movement drive module 400 can be configured to include a movement bar 420 coupled to one end of the constant temperature plate 350, a drive motor unit 410, and a conversion plate 430 that converts the rotational force of the drive motor unit 410 into a horizontal movement force of the movement bar 420, as described above.

[0148] As shown in FIG. 21, the PCR plate 200 is inserted into the nucleic acid extraction cartridge 100 and coupled in a structure where the flow paths are connected. The nucleic acid solution extracted from the nucleic acid extraction cartridge 100 is injected into the injection port h1, and then the injected extracted nucleic acid solution moves to the PCR plate 200 including one or more reaction wells containing a PCR mixture dry product containing at least one of a primer and a probe.

[0149] Thereafter, the first heating block 310 and the second heating block 320 of the temperature control module 300 of the present invention for forming the first temperature or the second temperature descend to the reaction well portion of the PCR plate 200.

[0150] In this case, the horizontal movement drive module 400 is configured to horizontally move both the thermostatic plate 350 and the temperature control module 300, and the PCR plate 200 is arranged in a structure inserted between the thermostatic plate 350 and the temperature control module 300.

[0151] When the heating blocks 310 and 320 horizontally moved through the horizontal movement drive module 400 press the PCR plate 200, the first region or the second region of the thermostatic plate 350 having a set temperature (the first temperature or the second temperature) corresponding to the temperature of the heating block 310 or 320 is arranged so as to naturally correspond.

[0152] That is, when the first region a1 of the thermostatic plate 350 horizontally moves and is arranged below the PCR plate 200, the first heating block (FIG. 2) 310 simultaneously horizontally moves so as to face the upper surface of the PCR plate 200, and then, the heating blocks 310 and 320 descend and come into contact with the upper surface of the PCR plate 200.

[0153] Also, when the second region a2 of the thermostatic plate 350 horizontally moves and is arranged below the PCR plate 200, the second heating block (FIG. 2) 320 operates corresponding to face the upper surface of the PCR plate 200 while horizontally moving simultaneously, and then, the heating blocks 310 and 320 descend and come into contact with the upper surface of the PCR plate 200.

[0154] In short, when performing temperature cycling on the PCR plate 200, to raise the temperature to the first temperature, the first heating block 310 horizontally moves so as to face the upper surface of the PCR plate 200, and the lower surface of the PCR plate 200 is driven so that after the first region of the thermostatic plate 350 horizontally moves, the first heating block 310 moves downward and comes into contact with and is pressed.

[0155] Also, to lower the temperature to the second temperature again, the upper surface of the PCR plate 200 moves horizontally so as to face the second heating block 320. On the lower surface of the PCR plate 200, after the second region of the constant temperature plate 350 moves horizontally together with the second heating block 320, the second heating block 320 moves downward while being brought into contact with and pressurized, and is driven so that heating and cooling are simultaneously performed on the upper and lower surfaces of the PCR plate 200.

[0156] In this way, by simultaneously performing contact and pressurization on the upper and lower surfaces of the PCR plate 200, the efficiency is twice that of the constant temperature plate 350 method maintained at a single temperature.

[0157] Hereinafter, with respect to the nucleic acid extraction cartridge 100 that embodies a PCR (polymerase chain reaction) premix or nucleic acid solution containing a nucleic acid extract in the PCR plate 200 in the present invention described above, it will be described with reference to FIGS. 24 to 32.

[0158] FIG. 24 is a perspective conceptual diagram of the nucleic acid extraction cartridge of the present invention, showing a structure in which the above-described PCR plate is inserted and coupled. FIG. 25 is a separated perspective view of FIG. 24, and FIG. 26 is a diagram showing the internal structure of the cartridge cover R1 in the structure of FIG. 25.

[0159] Referring to FIGS. 24 to 26, the nucleic acid extraction cartridge 100 according to the present invention includes a cartridge cover R1 having a plurality of partitioned structures accommodating portions 22, 23, 24, 25, 26 containing solutions necessary for DNA extraction, and a cartridge main body portion R2 provided with a reaction accommodating portion 11 that reacts or purifies the solution drawn from each of the accommodating portions 22, 23, 24, 25, 26 with a sample, and may be configured to include.

[0160] In this case, it is configured to include a piston 18 that injects the PCR premix or nucleic acid solution purified in the reaction accommodating portion 11 into the injection port h1 of the PCR plate 200 that is coupled in a structure inserted into the cartridge main body portion R2.

[0161] In the present invention, the cartridge cover R1 is in contact with one side surface of each reaction accommodating portion 11 of the cartridge main body portion R2, and includes a rubber portion 30 containing an elastically deformable material. The rubber portion 30 is made of an elastically deformable material and is composed of rubber, silicone, etc.

[0162] In the present invention, when the rubber portion 30 is assembled with the cartridge cover R1 and the cartridge main body portion R2, by sealing the space between the cartridge cover R1 and the cartridge main body portion R2, it can ride on the wall surface of the reaction accommodating portion 11 of the cartridge main body portion R2 to prevent the nucleic acid solutions from being mixed with each other.

[0163] The operation of the nucleic acid extraction cartridge of the present invention will be described with reference to FIGS. 25 to 32 as follows. FIG. 27 is a transparent perspective view of FIG. 24, showing the internal structure after bonding.

[0164] The nucleic acid extraction cartridge of the present invention is provided on the bottom surface of the main body portion R2 and is equipped with a rotary valve 19 in which a flow path 19-1 is formed inside. When this rotary valve is rotated, the spaces of the respective accommodating portions 11, 12, 13, 14, 15, 16, 17 of the cartridge main body portion R2 can be connected to the flow path of the rotary valve 19. After connecting the flow path to a specific accommodating portion and then operating the piston 18, it is designed to be able to collect the solution that has entered the accommodating portion and transfer substances to other accommodating portions or a PCR plate 200.

[0165] As shown in FIGS. 25 and 26, inside the cartridge cover R1, accommodating portions 22, 23, 24, 25, 26 containing the respective solutions necessary for DNA extraction are formed. Since the bottom surface of this accommodating portion is sealed with a film or the like, it is designed to be easily penetrated by the through needle of the main body accommodating portion. Also, five holes 21-1, 21-2, 27, 28, 29 are formed.

[0166] The binding buffer is contained in the first accommodation part 22 of the cartridge cover R1, the 1st washing buffer is contained in the second accommodation part 23, the 2nd washing buffer is contained in the third accommodation part 24, the 3rd washing buffer is contained in the fourth accommodation part 25, and the elution buffer is contained in the fifth accommodation part 26.

[0167] The PCR plate 200 is covered with a film made of a transparent plastic material (such as polyethylene, polypropylene, PET, etc.), and inside the reaction wells is a PCR dry matter mixture containing at least one side of the dried PCR primers and probes, which has the same structure as the one described with reference to FIG. 8.

[0168] The operation of the nucleic acid extraction cartridge of the present invention can proceed in the following order.

[0169] 1. Introduction of biological sample

[0170] With the cartridge main body R2, the cartridge cover R1, and the PCR plate 200 combined, they are mounted on the automatic equipment described later, and a biological sample (blood) is introduced into the first hole 21-1 shown in FIG. 25.

[0171] 2. Outflow of nucleic acid from cells and binding to beads

[0172] As shown in FIG. 30, through the rotation of the rotary valve 19 arranged at the lower part of the cartridge main body R2 and the action of the piston 18, the binding buffer in the first accommodation part 22 is introduced into the reaction accommodation part 11 and mixed with the biological sample and the beads of the magnetic tablet MT (magnetic beads coated with silica).

[0173] The magnetic tablet MT used in the present invention is mounted at the end of a through pipeline extending inside the reaction accommodating portion 11 of the cartridge main body R2, and functions such that the nucleic acid extracted from the cells contained in the biological sample is dissolved in the magnetic tablet and bound to the surface of the dispersed magnetic beads.

[0174] At this time, instead of the magnetic tablet, the magnetic beads may be suspended in the binding buffer and used.

[0175] Thereafter, when ultrasonic waves are applied while a sonication tip is inserted into the sealed second hole 21-2 of the cartridge main body R2, the ultrasonic waves are transmitted through the plastic, and the reaction solution is homogenized while the biological sample, the tablet, and the binding buffer are mixed. At this time, each biological tissue contained in the biological sample is also disrupted, so that the nucleic acid flows out, and the flowed-out nucleic acid is bound to the surface of the beads.

[0176] When a magnetic bar is inserted into the third hole 27 of the cartridge main body R2, the beads are fixed to the wall surface of the reaction accommodating portion, and the remaining reaction solution is transferred to the first accommodating portion through the rotation of the rotary valve and the action of the piston.

[0177] 3. Primary washing

[0178] Through the rotation of the rotary valve of the cartridge main body R2 shown in FIG. 27 and the action of the piston, the primary washing buffer in the second accommodating portion 23 is introduced into the reaction accommodating portion 11 and mixed with the beads to which the nucleic acid is bound.

[0179] Thereafter, when the magnetic bar is removed from the third hole 27 in FIG. 25 and ultrasonic waves are applied while the sonication tip is inserted into the second hole 21-2, primary washing is performed. By this primary washing, in addition to the nucleic acid, each substance non-specifically bound to the beads is washed.

[0180] While a magnetic bar is inserted into the third hole 27, the beads are fixed to the wall surface of the reaction accommodating section, and the primary cleaning liquid is transferred to the second accommodating section 23 through the rotation of the rotary valve and the action of the piston.

[0181] 4. Secondary washing

[0182] Through the rotation of the rotary valve and the action of the piston of the cartridge main body R2 shown in FIG. 27, the secondary cleaning buffer in the third accommodating section 24 is introduced into the reaction accommodating section 11 and mixed with the beads to which the nucleic acid is bound.

[0183] Thereafter, when the magnetic bar is removed from the third hole 27 in FIG. 25 and ultrasonic waves are applied while a vibrator is inserted into the second hole 21-2, secondary cleaning is performed. By this secondary cleaning, substances other than the nucleic acid that are non-specifically bound to the beads are washed away.

[0184] While a magnetic bar is inserted into the third hole 27, the beads are fixed to the wall surface of the reaction accommodating section, and the secondary cleaning liquid is transferred to the third accommodating section 24 through the rotation of the rotary valve and the action of the piston.

[0185] 5. Tertiary washing

[0186] Through the rotation of the rotary valve and the action of the piston of the cartridge main body R2 shown in FIG. 27, the tertiary cleaning buffer in the fourth accommodating section 25 is introduced into the reaction accommodating section 11 and mixed with the beads to which the nucleic acid is bound.

[0187] Thereafter, when the magnetic bar is removed from the third hole 27 in FIG. 25 and ultrasonic waves are applied while a vibrator is inserted into the second hole 21-2, tertiary cleaning is performed. By this tertiary cleaning, substances other than the nucleic acid that are non-specifically bound to the beads are washed away.

[0188] While a magnetic bar is inserted into the third hole 27, the beads are fixed to the wall surface of the reaction accommodating section, and the tertiary cleaning liquid is transferred to the fourth accommodating section 25 through the rotation of the rotary valve and the action of the piston.

[0189] 6. Nucleic acid elution

[0190] Through the rotation of the rotary valve of the cartridge main body R2 shown in FIG. 27 and the action of the piston, the elution buffer in the fifth storage unit 26 is introduced into the reaction storage unit 11 and mixed with the beads to which the nucleic acid is bound.

[0191] Thereafter, when the magnetic bar is removed from the third hole 27 in FIG. 25 and ultrasonic waves are applied while the oscillator is inserted into the second hole 21-2, the nucleic acid bound to the surface of the beads is dissolved in the elution buffer.

[0192] 7. Generation of PCR premix

[0193] While the magnetic bar is inserted into the third hole 27, the beads are fixed to the wall surface of the reaction storage unit. Through the rotation of the rotary valve and the selection of the small piston, the elution buffer in which the nucleic acid is dissolved is introduced into the sixth storage unit 17 and mixed with the PCR material (a mixture of polymerase, dNTP, etc.) in the sixth storage unit to generate a PCR preliminary mixture. The "PCR preliminary mixture" used in the present invention is defined and used as being embodied by the above substances.

[0194] 8. Transfer to PCR plate

[0195] Through the rotation of the rotary valve of the cartridge main body R2 shown in FIG. 27 and the action of the piston, the PCR preliminary mixture and the nucleic acid solution generated in the sixth storage unit are introduced into the PCR plate 200 and mixed with at least one side of the primers and probes in the PCR plate 200. Such an introduction process is injected into the injection port h1 while moving along the flow path Y of the rotary valve by the pressurization of the piston 18 as shown in FIG. 32.

[0196] Thereafter, while a heating rod is inserted into the fourth hole 29, the cover film at the inlet portion of the PCR reaction plate is pressurized and heated, whereby the PCR reaction plate is sealed.

[0197] 9. PCR reaction

[0198] Finally, the PCR plate 200 is in a state containing at least one of nucleic acids, polymerase, dNTP, primers, and probes extracted from a biological sample and the other buffer solution.

[0199] Therefore, the PCR reaction is performed by applying pressurized thermal application to this PCR plate 200 through the temperature control module of the present invention described above.

[0200] Figures 33 to 36 are diagrams showing the overall structure and arrangement configuration of the apparatus constituting the polymerase chain reaction system of the present invention described above.

[0201] The polymerase chain reaction apparatus according to an embodiment of the present invention can include a nucleic acid extraction cartridge assembly including a nucleic acid extraction cartridge 100 and a PCR plate 200.

[0202] As shown in Figure 35, when the polymerase chain reaction system according to an embodiment of the present invention is mounted inside the nucleic acid extraction cartridge 100, the above-described PCR plate 200 is placed on the side surface. The portion where the reaction wells corresponding to the body portion of the PCR plate 200 are present is exposed to the outside, and the above-described temperature control module 300 is arranged above it.

[0203] Figure 34 is an enlarged view of the coupling arrangement diagram of the main part of the present invention in Figure 33, Figure 35 is a vertical cross-sectional conceptual diagram of a part of Figure 34, presenting the arrangement of the main part configuration. Figure 36 is a side perspective cross-sectional conceptual diagram of Figure 35.

[0204] As shown in FIGS. 34 to 36, the PCR premix or nucleic acid solution containing the nucleic acid extracted from the inside of the nucleic acid extraction cartridge 100 according to the present invention is injected into the PCR plate 200 in which the reaction wells W are embodied. On the upper part of the PCR plate 200, a first pressurizing surface G1 corresponding to the surface of the reaction well W is embodied, and a first heating block 310 maintained at a temperature required for heat denaturation by a heating unit is arranged, and a second heating block 320 which can be embodied in a structure in which the region pressurized while moving horizontally is changed is arranged in proximity. Thus, the PCR premix and the nucleic acid solution injected into the reaction well W are directly heated by the heating block so as to conform to the first temperature (95° C.) required for heat denaturation and the second temperature (55° C.) required for annealing.

[0205] In addition, as shown in FIGS. 35 and 36, a constant temperature plate 350 is arranged below the PCR plate 200 so that the temperature of the PCR plate 200 is maintained at a constant temperature level.

[0206] A scanning module 500 is arranged below the constant temperature plate 350, and the light L irradiated by the light irradiation unit E1 reaches the PCR plate 200 through the light transmission part H of the constant temperature plate 350, and fluorescence detection is performed.

[0207] In the embodiment of the present invention, as described above, when the temperature is increased while the first heating block 310 at the first temperature (for example, 95° C.) is brought into close contact by the temperature control module 300, or when the second heating block 320 at the second temperature (for example, 55° C.) is brought into close contact, if the PCR plate 200 is maintained within a certain temperature range, the temperature control of the internal amplification reactant becomes much easier. Therefore, maintaining the temperature of the above-described constant temperature plate 350 constant at the second temperature acts as a very important factor for enhancing the reliability of the reaction. When the temperature of the PCR plate 200 is raised to 95° C., the heat transfer rate can be increased while bringing the first temperature block and the first constant temperature region into close contact, and the temperature of the PCR plate can reach 95° C. rapidly within 2 to 3 seconds.

[0208] When performing RT / PCR to detect an RNA target, a PCR premix or a PCR plate containing a dried RT-PCR reaction product is used. After closely attaching the PCR reaction plate to a low-temperature block while adjusting the temperature of the low-temperature block to the RT reaction temperature, the RT reaction can be maintained for the RT reaction time, and after performing the reverse transcription reaction, the PCR reaction can be performed.

[0209] Through the PCR reaction, the presence or absence or concentration of the amplified nucleic acid can be determined, and this information can be used for diagnosis. At this time, the presence or absence or concentration of the amplified nucleic acid can be determined using a normal nucleic acid detection method.

[0210] For example, a method using SYBR green, a DNA minor groove intercalating fluorescent dye which is a DNA intercalation dye, a method of scanning excitation light and corresponding fluorescence in various wavelength bands using a probe attached with various phosphors and quenchers, etc. are available, and it is not limited thereto.

[0211] Referring to FIGS. 37 to 44, in still another embodiment of the present invention, the body portion 210 of the PCR plate 200 includes a body frame portion 211, a blocking accommodation portion 213, and a connecting flow path portion 215. The body frame portion 211 includes one or more reaction wells W (W1... Wn).

[0212] The blocking accommodation portion 213 is formed in a concave shape in the body frame portion 211, is disposed corresponding to each reaction well W, and is formed in a circular shape so as to accommodate a circular blocking portion 240. The blocking accommodation portion 213 is formed in a concave shape corresponding to the size of the blocking portion 240 so as to accommodate the blocking portion 240.

[0213] The connecting flow path portion 215 connects the blocking accommodation portion 213 and each reaction well W. The nucleic acid solution can be supplied to the reaction well W through the connecting flow path portion 215.

[0214] The blocking part 240 is configured to include an elastically deformable material. The blocking part 240 is made of an elastically deformable material such as rubber or silicone. When injecting the nucleic acid solution into each reaction well W, since the blocking part 240 disposed in each blocking accommodation part 213 is made of rubber or the like and has a small self-weight, the resistance applied to each blocking accommodation part 213 is small.

[0215] Therefore, the nucleic acid solution is injected through one flow path part 230, and the same amount of nucleic acid solution is uniformly injected into each reaction well W.

[0216] Referring to FIGS. 43 and 44, a hole is formed below each blocking accommodation part 213, and the blocking part 240 contacts this hole due to its own weight. When the pressure in each reaction well W becomes higher than that on the side of the connecting flow path part 215, the blocking part 240 adheres closely to the hole of the blocking accommodation part 213, and can function as a check valve for preventing the backflow of the nucleic acid solution.

[0217] FIG. 45 is a diagram showing the test results for the outermost reaction wells W1 and W8 among the eight reaction wells W1 to W8 in the reaction well W. During the PCR reaction, in order to confirm the presence or absence of mixing of the nucleic acid solution between the reaction wells W, PCR was carried out after drying the reaction wells.

[0218] The reaction wells W were confirmed 5 minutes, 15 minutes, and 25 minutes after starting PCR. It was confirmed that the nucleic acid solution was mixed with the horizontal reaction wells W as time passed, and it was confirmed that all the reaction wells W1 to W8 were mixed 15 minutes later.

[0219] FIG. 46 is a diagram showing the test results for the outermost reaction wells W1 and W8 among the eight reaction wells W1 to W8 in the reaction well W. After drying the outermost reaction wells and then proceeding with PCR, it was confirmed that the solution between the reaction wells was not mixed by the check valve.

[0220] Referring to FIGS. 37 to 41, the nucleic acid extraction cartridge 100 is formed with holes 21-1, 21-2, 27, 28, and 29. Leakage prevention rubbers 40 are attached to each of the holes 21-1, 21-2, 27, 28, and 29 of the nucleic acid extraction cartridge 100. The leakage prevention rubber 40 is made of a material such as elastic rubber.

[0221] With this leakage prevention rubber 40, it is possible to prevent the nucleic acid solution remaining in the nucleic acid extraction cartridge 100 from leaking through each of the holes 21-1, 21-2, 27, 28, and 29 of the nucleic acid extraction cartridge 100.

[0222] Referring to FIG. 39, the leakage prevention rubber 40 is pierced by a semi-circular punch 50 to form a hole through which air can flow in.

[0223] Referring to FIG. 40, when the nucleic acid solution flows in, the leakage prevention rubber 40 is pushed by a pressing jig 60, and the atmospheric pressure can be smoothly inhaled through the hole of the leakage prevention rubber 40.

[0224] Referring to FIG. 41, when the nucleic acid extraction cartridge 100 is discarded, the leakage prevention rubber 40 can close the nucleic acid extraction cartridge 100 and prevent the leakage of the nucleic acid solution remaining inside the nucleic acid extraction cartridge 100.

[0225] Referring to FIG. 44, a sheet portion 250 is heat-sealed to the reaction well W. Each reaction well W is heat-sealed by the sheet portion 250 to maximize the volume of each reaction well W, and heat transfer can proceed rapidly within the reaction well W, and the reaction of the nucleic acid solution can be carried out rapidly.

[0226] The present invention has been described with reference to one embodiment shown in the drawings, but this is merely illustrative, and those having ordinary knowledge in the art to which the technology pertains will understand that various modifications and equivalent other embodiments will be possible hereafter. Therefore, the true technical protection scope of the present invention should be defined by the following claims.

Claims

1. A body part having one or more reaction wells; An insertion part extending from the body part, inserted into a nucleic acid extraction cartridge, and having an injection port into which a nucleic acid solution is injected; A flow path part for making the nucleic acid solution flow from the injection port to the reaction wells; and A blocking part attached to the body part for blocking backflow of the nucleic acid solution from the reaction wells to the flow path part side; A PCR plate characterized by including.

2. The body part is A body frame part having one or more of the reaction wells; A blocking accommodation part formed in a concave shape in the body frame part for accommodating the blocking part; A connecting flow path part connecting the blocking accommodation part and the reaction wells; A storage part formed in a concave shape in the body part, communicating with the flow path part, and receiving the supply of the nucleic acid solution; and A flow path guide part communicating with the storage part for guiding the nucleic acid solution to flow toward the blocking part side; The PCR plate according to claim 1, characterized by including.

3. The connecting flow path part includes a bottleneck part formed with a narrow width from the blocking part toward the reaction well side; The PCR plate according to claim 2, characterized by this.

4. The blocking accommodation part is divided into a plurality of parts by a partition part formed along the length direction of the storage part, The PCR plate according to claim 2, characterized in that the blocking part is attached to each of the divided blocking accommodation parts.

5. The blocking part is configured to include an elastically deformable material; The PCR plate according to claim 1, characterized by this.

6. The body part is A body frame part having one or more of the reaction wells; A blocking accommodation part formed in a concave shape in the body frame part, arranged corresponding to each of the reaction wells, and formed in a circular shape so as to accommodate the circular blocking part; and A connecting flow path part connecting the blocking accommodation part and the reaction wells; The PCR plate according to claim 1, characterized by including.

7. The blocking part is configured to include an elastically deformable material; The PCR plate according to claim 6, characterized by this.

8. A sheet part is thermally fused to the reaction wells; The PCR plate according to claim 6, characterized by this.

9. A cartridge cover having accommodation parts with a plurality of partition structures containing solutions necessary for DNA extraction; and A cartridge main body part that is coupled to the lid part by an insertion structure and is provided with a reaction accommodation part that reacts or purifies a solution drawn from the accommodation part with a sample; A nucleic acid extraction cartridge characterized by including.

10. The cartridge cover is in contact with one side surface of the accommodation part of the cartridge main body part, and includes a rubber part including an elastically deformable material. The nucleic acid extraction cartridge according to claim 9.

11. The nucleic acid extraction cartridge according to claim 9, wherein a liquid leakage prevention rubber including an elastically deformable material is attached to the nucleic acid extraction cartridge to prevent leakage of the nucleic acid solution.

12. A nucleic acid extraction cartridge; and a PCR plate that receives a nucleic acid solution from the nucleic acid extraction cartridge and accommodates the nucleic acid solution in a reaction well in which a PCR mixture dry product is accommodated, The nucleic acid extraction cartridge is A cartridge cover having an accommodation part with a plurality of partition structures containing solutions necessary for DNA extraction; and A cartridge main body part that is coupled to the lid part by an insertion structure and is provided with a reaction accommodation part that reacts or purifies a solution drawn from the accommodation part with a sample; including The PCR plate is A body part having one or more of the reaction wells; An insertion part extending from the body part, inserted into the nucleic acid extraction cartridge, and having an injection port into which the nucleic acid solution is injected; A flow path part that enables the nucleic acid solution to flow from the injection port to the reaction well; and A nucleic acid extraction cartridge assembly, characterized by including a blocking part that is attached to the body part and blocks the backflow of the nucleic acid solution from the reaction well to the flow path part side.

13. The cartridge cover is in contact with one side surface of the accommodation part of the cartridge main body part, and includes a rubber part including an elastically deformable material. The nucleic acid extraction cartridge assembly according to claim 12.

14. The nucleic acid extraction cartridge according to claim 12, wherein a liquid leakage prevention rubber including an elastically deformable material is attached to the nucleic acid extraction cartridge to prevent leakage of the nucleic acid solution.

15. A nucleic acid extraction cartridge; and a PCR plate that receives a nucleic acid solution with the nucleic acid extraction cartridge and accommodates the nucleic acid solution in a reaction well in which a PCR mixture dry product is accommodated, The nucleic acid extraction cartridge is A cartridge cover having an accommodation part with a plurality of partition structures containing solutions necessary for DNA extraction; and A cartridge main body part provided with a reaction accommodating part that is coupled to the lid part and reacts or purifies a solution drawn from the accommodating part with a sample; The PCR plate is A body part having one or more of the reaction wells; An insertion part extending from the body part, inserted into the nucleic acid extraction cartridge, and having an injection port into which the nucleic acid solution is injected; A flow path part that enables the nucleic acid solution to flow from the injection port to the reaction wells; and A blocking part attached to the body part and blocking backflow of the nucleic acid solution from the reaction wells to the flow path part side. The polymerase chain reaction device is characterized by including the above.

16. The cartridge cover is in contact with one side surface of the accommodating part of the cartridge main body part and includes a rubber part containing an elastically deformable material. The polymerase chain reaction device according to claim 15 is characterized by this.

17. The nucleic acid extraction cartridge is characterized in that a liquid leakage prevention rubber containing an elastic material and preventing leakage of the nucleic acid solution is attached thereto. The polymerase chain reaction device according to claim 15 is characterized by this.

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