PCR plate, nucleic acid extraction cartridge, and polymerization enzyme chain reaction apparatus 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 nucleic acids in real time the amplified product.

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 the symptoms of a disease at once within a short time based on the symptoms of the disease such as cough, diarrhea, high fever, genital abnormalities, etc., the causative pathogen is confirmed, and the on-site diagnosis based on symptoms that prescribes the optimal antibiotic and therapeutic agent is the core new technology of future precision medicine, and many studies are being developed and evolving. Such on-site diagnostic technology has the advantage of being able to be 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 is attracting attention as a core technology of future medicine that can utilize these technologies to accurately identify the cause in 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, current molecular diagnostic systems require 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] A representative molecular diagnostic method is the polymerase chain reaction (PCR). Invented by Kary Mullis in 1985, PCR has been widely used in molecular biology and molecular diagnostics because it allows for the rapid and easy amplification of specific DNA. PCR / RT-PCR allows for the confirmation of the presence of specific DNA / RNA in a biological sample, making it widely used in the diagnosis of pathogenic microbial infections such as viruses. This PCR / RT-PCR technique has evolved into Realtime Quantitative PCR (RT-PCR), which allows for simultaneous confirmation of results, simplifying the testing process and significantly reducing testing time. Furthermore, it accurately quantifies the number of pathogens, making it a standard diagnostic method for monitoring the effectiveness of treatments for viruses such as HIV, HCV, and HBV. Additionally, PCR / RT-PCR technology allows for the examination of gene expression patterns and gene mutations associated with specific diseases, making it a crucial technique for disease diagnosis.

[0005] To perform PCR in this manner, a nucleic acid extraction step is necessary to remove substances that inhibit 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 both the biological sample and the nucleic acid extraction procedure. When performed manually, problems such as contamination due to worker error can occur, so molecular diagnostics are now performed using almost entirely automated nucleic acid extraction equipment.

[0006] Since real-time quantitative PCR equipment is required to detect PCR reactions and reaction products, molecular diagnostics were previously mainly performed in large hospitals and specialized clinical testing laboratories.

[0007] Through recent research and development, a variety of automated systems and devices have been developed that automate the entire process of nucleic acid extraction, PCR reaction, and reaction product detection, making PCR easily accessible even without specialized skills.

[0008] However, existing devices have problems such as being excessively expensive, requiring long processing times, and making it difficult to perform a variety of tests at once.

[0009] To explain the basic principle of PCR, a DNA double helix is ​​heated to 95°C to separate it into single strands. Then, the reaction solution is cooled to the annealing temperature, and complementary primers in the PCR reaction solution are selectively mixed to both ends of the region to be amplified. DNA polymerase then sequentially links the four types of nucleotide triphosphates (A, G, T, and C) complementary to each single strand, repeating the reaction to create a double helix. PCR is experimentally performed by repeating the heating and cooling of the PCR reaction solution 30 to 45 times (n) to amplify a specific DNA double helix. n This is a reaction that geometrically amplifies a single molecule. RT-PCR has been extended to detect RNA by synthesizing cDNA through reverse transcription and then making it amplified via PCR.

[0010] To fully utilize PCR in molecular diagnostics, the newly developed real-time quantitative PCR principle is as follows: For quantitative analysis of DNA amplified using the PCR reaction, a substance that emits fluorescence in proportion to the amount of DNA is added to the PCR reaction solution. Fluorescence is then measured for each cycle, and the cycle in which a critical fluorescence value is detected is identified. From this, the initial concentration of the target nucleic acid is quantitatively measured.

[0011] Since the invention of PCR, while a variety of application technologies have been developed, the base sequences of numerous pathogens and disease-related genes have been known through genome projects. Molecular diagnostics that amplify these disease-related DNA / RNA base sequences and diagnose them qualitatively and quantitatively have rapidly developed. Existing PCR processes take about two hours to cycle through temperature, 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] To perform a PCR reaction quickly, the temperature of the reaction solution must be rapidly changed. Furthermore, in order to amplify only the desired target with an accurate PCR reaction, each primer must be designed to specifically attach to the desired target, and the annealing temperature must be precisely controlled in a PCR temperature cycling reaction.

[0013] For this purpose, micro-PCR reaction vessels were developed that have the smallest possible heat capacity and efficient heat transfer compared to the 0.2 ml and 0.5 ml reactors commonly used in existing laboratories. Each of these micro-reactors uses less reaction solution and has a large surface area, allowing for rapid heat transfer and quick heating and cooling. A large surface area was maintained by placing 10 μl of PCR solution into thin reaction grooves (40 μm to 80 μm) formed on a silicon wafer in a 17 × 15 mm area and covering it with a glass plate (>100 mm). 2 ( / 10μl), it was not possible to shorten the time to approximately 3 minutes per cycle using the existing Peltier thermal block method (Clin. Chem. 40 / 9, 1815-1818 (1994)).

[0014] In early PCR reactors, a method was developed to rapidly circulate the heat of the PCR reactor by repeatedly immersing it in high-temperature and low-temperature water baths (Turbo Thermalcycler, Bioneer Corp., Daejeon). This type of PCR equipment, which circulates the reactor between areas with different temperatures, is a spatial transfer method and has the advantage of enabling rapid and accurate PCR reactions by immersing the reactor in a constant-temperature water bath where the temperature is precisely maintained beforehand. However, because it requires many constant-temperature water baths, the equipment is large and difficult to maintain. Therefore, PCR equipment that employs a time-difference temperature circulation method, where the temperature is changed over time using a fixed block and a Peltier element or similar device, has become the mainstream.

[0015] PCR methods using microchannels have also been developed into spatial transfer temperature circulation methods and time-difference temperature circulation methods. Spatial transfer temperature circulation methods can be broadly divided into open-type methods where the solution flows continuously in a FIFO (First-In-First-Out) manner, and closed-type methods where the solution repeatedly moves between different temperature sections. The open-type method was developed in 1994 by Nakano et al. in which a capillary tube is wrapped around a cylindrical block with compartments of different temperatures, and the PCR solution is continuously flowed through it (Biosci. Biotech. Biochem., 58(2), 349-352, 1994). In 1998, using a microchannel PCR system in the form of a microchannel, Kopp et al. confirmed that PCR could be carried out by passing 10 μl of solution through only 20 cycles at 4.5-second cycles (Science 280 1046-1048, 1998).

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

[0017] Conventional Korean Patent No. 10-2105558 discloses a high-speed polymerase chain reaction (PCR) analysis plate. However, when using the disclosed analysis plate, a problem may arise in which the amplified products between each reaction well are mixed during the polymerase chain reaction process, reducing the accuracy of the experiment. To solve this problem, there is a need for an improved PCR plate that can rapidly perform quantitative and qualitative analysis of a large number of targets. [Overview of the Initiative] [Problems that the invention aims to solve]

[0018] The present invention has been made to solve the above-mentioned problems, and the object of the present invention is to provide a PCR plate, a nucleic acid extraction cartridge, and a polymerization enzyme chain reaction apparatus containing the same that can perform fully automated target nucleic acid detection through nucleic acid extraction from a biological sample, PCR reaction, excitation light of various wavelengths and corresponding fluorescence scanning, can examine a large number of targets in a single operation, is easy to use, and in particular can obtain accurate results in a short time.

[0019] Furthermore, an object of the present invention is to provide a PCR plate, a nucleic acid extraction cartridge, and a polymerization enzyme chain reaction apparatus containing the same, which enable rapid and accurate PCR by rapidly and repeatedly applying the temperature required for the thermal denaturation process and the precise temperature required for the binding process to the reactants during the temperature control process necessary for the PCR process, thereby maximizing the reliability of the reaction. [Means for solving the problem]

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

[0021] In the present invention, the body portion may include: a body frame portion having one or more reaction wells; a blocking housing portion formed concavely in the body frame portion and housing the blocking portion; a connecting channel portion connecting the blocking housing portion and the reaction wells; a storage portion formed concavely in the body portion, communicating with the channel portion and receiving the nucleic acid solution; and a channel guide portion communicating with the storage portion and guiding the nucleic acid solution to flow toward the blocking portion.

[0022] In the present invention, the connecting channel portion may include a bottle neck portion that is narrower in width from the blocking portion toward the reaction well.

[0023] In the present invention, the blocking accommodation part may be divided into a plurality of parts by a partition 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 a material that can be elastically deformed.

[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 that is formed in a concave shape in the body frame part, is arranged corresponding to each of the reaction wells, and is formed in a circular shape so as to accommodate the circular blocking part; and a connecting flow path part that connects the blocking accommodation part and the reaction well.

[0026] In the present invention, the blocking part may be configured to include a material that can be elastically deformed.

[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 that is coupled to the lid part by an insertion structure and is 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 a material that can be elastically deformed.

[0030] 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.

[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 places it in a reaction well containing a dried PCR mixture; wherein the nucleic acid extraction cartridge includes a cartridge cover having a plurality of partitioned containment sections containing a solution necessary for DNA extraction; and a cartridge body connected to the lid by an insertion structure and provided with a reaction containment section for reacting or purifying the solution drawn in from the containment section with a sample; wherein the PCR plate may include a body section having one or more reaction wells; an insertion section extending from the body section and inserted into the nucleic acid extraction cartridge, having an injection port into which the nucleic acid solution is injected; a flow channel section that allows the nucleic acid solution to flow from the injection port into the reaction well; and a blocking section attached to the body section that blocks the backflow of the nucleic acid solution from the reaction well to the flow channel section.

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

[0033] In the present invention, the nucleic acid extraction cartridge may include an elastic material and be fitted with a leak-proof rubber to prevent leakage of the nucleic acid solution.

[0034] The polymerization enzyme chain reaction apparatus 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 places it in a reaction well containing a dried PCR mixture; wherein the nucleic acid extraction cartridge includes a cartridge cover having a plurality of partitioned containment sections containing a solution necessary for DNA extraction; and a cartridge body connected to the lid by an insertion structure, which is provided with a reaction containment section for reacting or purifying the solution drawn in from the containment section with a sample; and the PCR plate may include a body section having one or more reaction wells; an insertion section extending from the body section and inserted into the nucleic acid extraction cartridge, which has an injection port into which the nucleic acid solution is injected; a flow channel section that allows the nucleic acid solution to flow from the injection port into the reaction well; and a blocking section attached to the body section that blocks the backflow of the nucleic acid solution from the reaction well to the flow channel section.

[0035] In the present invention, the cartridge cover may include a rubber portion that is in contact with one side of the housing portion of the cartridge body and is made of an elastically deformable material.

[0036] In the present invention, the nucleic acid extraction cartridge may include an elastic material and be fitted with a leak-proof rubber to prevent leakage of the nucleic acid solution. [Effects of the Invention]

[0037] According to the present invention, nucleic acid extraction from biological samples, PCR reactions, and real-time detection of reaction products through excitation light of various wavelengths and corresponding fluorescence scanning can be automated, allowing for various tests to be performed in a single operation, and the system is easy to use and provides accurate results in a short time.

[0038] Furthermore, in a PCR plate, it is possible to prevent the dried PCR mixture from flowing back out of the reaction well, thus preventing contamination of adjacent reaction wells by mixing with the dried PCR mixture.

[0039] Furthermore, according to the present invention, in the temperature control required for the PCR process, the precise temperature required for the thermal denaturation step and binding can be rapidly applied to the reactants in real time and simultaneously, thereby maximizing the reliability of the reaction and enabling accurate PCR.

[0040] In other words, the present invention eliminates the problems that arise when conventional temperature control methods, which increase the temperature while moving the reaction solution, cannot increase the temperature uniformly, which is detrimental to the PCR reaction, and when methods that cause a sequential temperature increase while the reaction solution is moving cannot simultaneously equalize the temperature throughout the entire reactant, thus increasing the likelihood of other reactions occurring. Instead, the present invention maintains a constant temperature within the temperature range set in the heating block and increases the temperature by directly pressurizing the entire reaction solution, thereby very efficiently achieving the temperature increase necessary for the PCR reaction.

[0041] Furthermore, to minimize the time delay during the process of changing the temperature from high to low, block-shaped heating block structures are arranged side by side with some space between them. When pressurizing a PCR plate, the position of the heating blocks is changed, and pressurization is performed in real time by heating blocks that have individually different temperatures. This revolutionary solution eliminates the problems caused by the time delay required during the temperature change process.

[0042] Furthermore, by incorporating the PCR plate into an insertable structure within the nucleic acid extraction cartridge, the nucleic acid extraction cartridge can be used commonly, allowing for the storage of PCR plates used in various test kits in a small space, and enabling the insertion and use of the appropriate PCR plate as needed during testing. Up to six fluorescence values ​​can be analyzed in a single reaction well within the PCR plate, and the number of reaction wells in the PCR plate can be increased to eight if necessary. This allows for the amplification and detection of all pathogenic bacteria that may be present in the patient's biological sample associated with symptoms, thus enabling the provision of symptom-based multimolecular diagnostic testing.

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

[0044] Furthermore, in realizing the mobile constant temperature plate structure, the reliability of the product's configuration and movement is ensured by using a sliding tape in the drive mechanism, and the inspection time can be shortened by adopting a method that simultaneously heats the plate inside the target on both the upper and lower surfaces. [Brief explanation of the drawing]

[0045] [Figure 1] This is a block diagram showing the main components of a polymerization enzyme chain reaction apparatus according to an embodiment of the present invention. [Figure 2] This figure illustrates the structure of a temperature control module in an embodiment of the present invention. [Figure 3] This figure illustrates the structure of a temperature control module in an embodiment of the present invention. [Figure 4] This figure illustrates the structure of a temperature control module in an embodiment of the present invention. [Figure 5] This figure illustrates the structure of a temperature control module in an embodiment of the present invention. [Figure 6] This figure illustrates the structure of a temperature control module in an embodiment of the present invention. [Figure 7] This figure illustrates the structure of a temperature control module in an embodiment of the present invention. [Figure 8] This figure shows one example of a PCR plate applied to the embodiments of the present invention. [Figure 9] This figure shows one example of a PCR plate applied to the embodiments of the present invention. [Figure 10] This figure shows one example of a PCR plate applied to the embodiments of the present invention. [Figure 11] This figure shows one example of a PCR plate applied to the embodiments of the present invention. [Figure 12] This figure shows one example of a PCR plate applied to the embodiments of the present invention. [Figure 13] This figure shows one example of a PCR plate applied to the embodiments of the present invention. [Figure 14] This figure shows another example of a PCR plate applied to the embodiments of the present invention. [Figure 15] This figure shows another example of a PCR plate applied to the embodiments of the present invention. [Figure 16] This figure shows another example of a PCR plate applied to the embodiments of the present invention. [Figure 17] This figure shows another example of a PCR plate applied to the embodiments of the present invention. [Figure 18] This figure shows another example of a PCR plate applied to the embodiments of the present invention. [Figure 19] This figure shows another example of a PCR plate applied to the embodiments of the present invention. [Figure 20] This is a conceptual diagram illustrating the structure and operation of a constant temperature plate and a horizontal movement drive module applied to embodiments of the present invention. [Figure 21] This is a conceptual diagram illustrating the structure and operation of a constant temperature plate and a horizontal movement drive module applied to embodiments of the present invention. [Figure 22] This is a conceptual diagram illustrating the structure and operation of a constant temperature plate and a horizontal movement drive module applied to embodiments of the present invention. [Figure 23] This is a conceptual diagram illustrating the structure and operation of a constant temperature plate and a horizontal movement drive module applied to embodiments of the present invention. [Figure 24]This 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 PCR plate described above is inserted and bound. [Figure 25] Figure 24 is a separated perspective view. [Figure 26] This figure shows the internal structure of the cartridge cover in the structure shown in Figure 24. [Figure 27] This is a perspective view showing the bonding state of the structure in Figure 24. [Figure 28] This is an assembly perspective view of another embodiment of the nucleic acid extraction cartridge of the present invention. [Figure 29] This is a cross-sectional view of another embodiment of the nucleic acid extraction cartridge of the present invention. [Figure 30] This figure shows the lower operating state of the cartridge structure according to an embodiment of the present invention. [Figure 31] This figure shows the lower operating state of the cartridge structure according to an embodiment of the present invention. [Figure 32] This figure shows the lower operating state of the cartridge structure according to an embodiment of the present invention. [Figure 33] This figure shows the overall structure and arrangement of the apparatus constituting the polymerization enzyme chain reaction apparatus, which is an embodiment of the present invention described above. [Figure 34] Figure 33 is an enlarged view of the combined arrangement of the main parts of the embodiment of the present invention. [Figure 35] This is a conceptual vertical cross-sectional view of the section shown in Figure 34, illustrating the arrangement of the main components. [Figure 36] Figure 35 is a side perspective cross-sectional view. [Figure 37] This is a perspective conceptual diagram of a nucleic acid extraction cartridge according to another embodiment of the present invention. [Figure 38] This is a cross-sectional view shown in Figure 37. [Figure 39] Figure 38 shows how to punch out leak-proof rubber using a punch. [Figure 40] Figure 39 shows the process of applying pressure to the leak-preventing rubber using a pressing jig. [Figure 41]This diagram shows how the leak-proof rubber prevents residual liquid leakage. [Figure 42] This figure shows yet another example of a PCR plate. [Figure 43] This is a cross-sectional view of Figure 42. [Figure 44] This diagram shows the sheet attached to the PCR plate. [Figure 45] This diagram shows a test to check whether the solutions in the reaction wells are mixed during PCR. [Figure 46] This figure shows a mixing test of reaction wells in a PCR plate. [Modes for carrying out the invention]

[0046] Hereinafter, an embodiment of the PCR plate, nucleic acid extraction cartridge, and polymerization enzyme chain reaction apparatus containing the same according to the present invention will be described with reference to the attached drawings. In this process, the thickness of the lines and the size of the components shown in the drawings may be exaggerated for clarity and convenience of explanation.

[0047] Furthermore, the terms described later are defined in consideration of the functions of the present invention, and these may change depending on the intent or conventions of the user and operator. Therefore, these terms should be defined based on the content throughout this specification.

[0048] Figure 1 is a block diagram showing the main components of a polymerization enzyme chain reaction apparatus according to an embodiment of the present invention; Figures 2 to 7 are diagrams illustrating the structure of the temperature control module in an embodiment of the present invention; Figures 8 to 13 show one embodiment of the PCR plate applied to an embodiment of the present invention; Figures 14 to 19 show other embodiments of the PCR plate applied to an embodiment of the present invention; Figures 20 to 23 are conceptual diagrams illustrating the structure and operation of the constant temperature plate and horizontal movement drive module applied to an embodiment of the present invention; Figure 24 is a perspective conceptual diagram of the nucleic acid extraction cartridge in an embodiment of the present invention, showing the structure in which the above-mentioned PCR plate is inserted and bonded; Figure 25 is a separated perspective view of Figure 24; Figure 26 shows the internal structure of the cartridge cover in the structure of Figure 24; Figure 27 is a perspective view showing the bonded state of the structure of Figure 24; Figure 28 is an assembled perspective view of another embodiment of the nucleic acid extraction cartridge in an embodiment of the present invention; Figure 29 is a cross-sectional view of another embodiment of the nucleic acid extraction cartridge in an embodiment of the present invention; Figures 30 to 32 show the cartridge structure of an embodiment of the present invention. Figure 33 shows the overall structure and arrangement of the apparatus constituting the polymerization enzyme chain reaction apparatus, which is an embodiment of the present invention described above. Figure 34 is an enlarged view of the coupling arrangement of the main parts of the embodiment of the present invention in Figure 33. Figure 35 is a conceptual vertical cross-sectional view of the part of Figure 34, showing the arrangement of the main components. Figure 36 is a side perspective cross-sectional view of Figure 35. Figure 37 is a conceptual perspective view of a nucleic acid extraction cartridge according to another embodiment of the present invention. Figure 38 is a cross-sectional view of Figure 37. Figure 39 shows the leakage prevention rubber being punched in Figure 38. Figure 40 shows the pressurization of the leak-proof rubber with a pressing jig in Figure 39, Figure 41 shows the leak-proof rubber preventing residual liquid leakage, Figure 42 shows yet another embodiment of the PCR plate, Figure 43 is a cross-sectional view of Figure 42, Figure 44 shows the sheet mounted on the PCR plate, Figure 45 shows a test to confirm whether the solutions between the reaction wells are mixed during PCR, and Figure 46 shows a mixing test of the reaction wells in the PCR plate.

[0049] Referring to Figure 1, the polymerase chain reaction apparatus according to an embodiment of the present invention may be equipped with a temperature control module, which is embodied in a heating block structure that contacts the PCR reaction plate and applies a specific temperature, enabling accurate PCR by applying the temperature required for the denaturation process and the precise temperature required for the annealing process to the reactants simultaneously in real time without any time difference during the temperature control process required for the PCR process, thereby maximizing the reliability of the reaction.

[0050] The temperature control module according to the present invention minimizes the time delay required to change the temperature from a first temperature to a relatively lower second temperature, or vice versa, by changing the positions of the heating blocks set for the first and second temperatures, respectively. This allows for real-time pressurization of the PCR reaction plate and revolutionizes the problem caused by the time delay required during the temperature change process.

[0051] Furthermore, the present invention can be further embodied by providing a constant-temperature plate structure positioned below the PCR plate and operating in a sliding manner. In this case, the constant-temperature plate structure maintains the temperature of the PCR plate at a first or second temperature, thereby minimizing the time required to apply temperature change conditions and maximizing the reaction rate.

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

[0053] The present invention, with the configuration described above, provides the convenience of allowing even non-experts to freely extract nucleic acids by adding the desired sample through the nucleic acid extraction cartridge 100. At the same time, in the temperature circulation necessary for amplification applied to the PCR plate 200, rapid and precise temperature control is possible using the heating block 310, 320 structures, which can directly apply the target temperature to the reaction solution in the PCR plate 200 through compression with a thin film.

[0054] Furthermore, the scanning module enables the provision of a polymerase chain reaction (PCR) apparatus that embodies a single system capable of performing real-time detection of reactants at the bottom of the PCR plate 200 through excitation light of various wavelengths and corresponding fluorescence scanning.

[0055] Figures 2 to 7 are diagrams illustrating the structure of the temperature control module 300 in the present invention.

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

[0057] Referring to Figures 2 and 3, the temperature control module 300 has the function of controlling the temperature of the PCR plate 200, which receives and contains a PCR (polymerase chain reaction) preliminary mixture or nucleic acid solution from the nucleic acid extraction cartridge 100, while extracting nucleic acids from a biological sample and mixing them with polymerization enzymes.

[0058] The temperature control module 300 may include a first heating block 310 on which a first pressure surface G1 corresponding to the surface of the reaction well W on the PCR plate 200 is provided, and which is maintained by a heating unit at a temperature set within the range of the temperature required for thermal denaturation (hereinafter, "first temperature"). In addition, it may be configured to include a second heating block 320 positioned at a distance from the first heating block 310, which is provided with a second pressure surface G2 corresponding to the surface of the reaction well W, and which is maintained by a heating unit at a temperature set within the range of the temperature required for annealing (hereinafter, "second temperature"). In particular, the structures of the first heating block 310 and the second heating block 320 are designed to be capable of horizontal and vertical movement.

[0059] In one embodiment, the first heating block 310 and the second heating block 320 have a three-dimensional structure and can have a flat pressure surface on their 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] The first heating block 310 and the second heating block 320 are arranged opposite each other in a mutually opposing structure, as shown in Figures 2 and 3. The upper surface of the two heating blocks is generally embodied in a flat structure for pressurizing, and the upper part may be provided in a rectangular parallelepiped three-dimensional structure. The embodiment of a rectangular parallelepiped three-dimensional structure is just one embodiment; any three-dimensional shape that has a flat pressurizing surface for pressurizing can be said to be included in the essence of the present invention.

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

[0062] In other words, the first temperature of the first heating block 310 may be set in the range of 94°C to 96°C, which corresponds to the temperature applied to the thermal denaturation step for separating double helix DNA (including DNA extracted from a biological sample), and in one example of the present invention, it may be maintained at 95°C.

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

[0064] The first heating block 310 and the second heating block 320 are not designed to contain water or a heat-transmitting fluid, but rather feature a metal body with high heat capacity and good heat transfer efficiency, allowing them to be constantly maintained at a 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 regulation to maintain a constant temperature.

[0065] In other words, when a PCR premixture or nucleic acid solution is injected into the PCR plate 200, the first heating block 310 moves horizontally to become adjacent to the surface of the PCR plate 200 when the first temperature needs to be applied. That is, since the first pressure surface G is a flat plate structure, the entire surface of the PCR plate 200 can be heated simultaneously at the same temperature and pressure, enabling uniform temperature transfer to the entire sample.

[0066] Furthermore, when it is necessary to apply the second temperature required for the binding stage, the second heating block 320 moves horizontally to position itself above the PCR plate 200, enabling simultaneous heating of the entire surface of the PCR plate 200 at the same temperature and pressure.

[0067] In other words, since there is no need to take separate time to prepare the set temperature reaction, and the system is driven in a way that allows the entire surface of the PCR plate 200 to be heated simultaneously at the same temperature and pressure with simple horizontal movement, it becomes possible to produce a faster and more precise PCR reaction compared to existing methods for controlling the set temperature.

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

[0069] Since it is important that the second heating block 320 can be maintained at a relatively second temperature, for example, an annealing temperature of 55°C, it can be equipped with a radiating cooling pattern on its upper part that minimizes thermal interference with the first heating block 310 and allows excess heat to be easily dissipated by a cooling fan unit. As an example of this, the present invention may further include a temperature control pattern section 321 in the second heating block 320 that is realized on the side surface of the second pressurized surface G2. The temperature control pattern section 321 has a structure in which many protruding patterns are realized on its upper part, which increases the heat dissipation efficiency while increasing the surface area in contact with air, thus making it advantageous to maintain a constant low temperature.

[0070] Unlike methods that involve moving the reaction sample embodying the PCR reaction or moving it to other heating regions over time, the present invention applies a heating block structure that allows the reaction sample to be fixed in place and simultaneously raise the temperature uniformly and evenly over the top, thereby enabling precise transfer of the first and second temperatures.

[0071] In addition, the first heating block 310 and the second heating block 320 in the present invention can be linked with a drive module 330 that implements horizontal or vertical movement. 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] In other words, the first heating block 310 and the second heating block 320 in the present invention are arranged apart from each other and move up and down while intersecting each other through the operation of the drive module 330. Furthermore, elastic members S1 and S2 are further included below the guide members 331 and 332, so that when the heating blocks 310 and 320 pressurize the PCR plate 200, a buffering effect can be provided by applying appropriate elastic force (see Figure 5).

[0073] Furthermore, the polymerization enzyme chain reaction apparatus according to the embodiment of the present invention may further include a constant temperature plate 350 that is linked to a temperature control module 300.

[0074] As shown in Figures 2 and 3, the constant temperature plate 350 is positioned below the heating block 310 and 320 structures that constitute the temperature control module 300. After the PCR plate 200 is inserted, when the first heating block 310 and second heating block 320 of the temperature control module 300 pressurize the PCR plate 200 through horizontal and vertical movement, the constant temperature plate 350 functions to maintain the same temperature as the first heating block 310 and second heating block 320.

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

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

[0077] As shown in Figures 2 and 3, the horizontal movement drive module 400 may be configured to include a moving bar 420 and a drive motor unit 410 connected to one end of the constant temperature plate 350, 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 downwards from the temperature control module 300 described above. In particular, the constant temperature plate 350 according to an embodiment of the present invention may be implemented with a structure divided 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 the embodiment of the present invention, the temperature control module 300 and the constant temperature plate 350 can be realized as an integrated unit with respect to guide members 331 and 332.

[0080] In other words, when the horizontal movement drive module 400 is driven, the temperature control module 300, which includes the constant temperature plate 350 and the heating blocks 310 and 320, can be moved together.

[0081] In this case, the constant temperature plate 350 includes a first region heated to a first temperature and a second region heated to a second temperature separate from the first region. The first pressurizing surface G1 of the first heating block 310 is positioned corresponding to the upper part of the first region. The PCR plate 200 can be placed between the constant temperature plate 350 and the heating blocks 310 and 320 on its upper and lower surfaces, and pressurized at the same temperature simultaneously.

[0082] In other words, in the embodiment of the present invention, the constant temperature plate 350 is divided into regions having a gradient between a first temperature and a second temperature, and when the heating block is pressurized through the horizontal movement drive module 400, the region having a set temperature (first temperature or second temperature) corresponding to the temperature of the heating blocks 310 and 320 moves horizontally in a sliding structure, and by simultaneously contacting and pressurizing the upper and lower surfaces of the PCR plate 200, twice the efficiency is achieved compared to the constant temperature plate 350 method which maintains a single temperature.

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

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

[0085] Referring to Figures 2 to 5, the drive module 330 moves the first heating block 310 and the second heating block 320 up and down, and simultaneously, the horizontal movement drive module 400 moves the first heating block 310 and the second heating block 320 horizontally, 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 pressure surface G1 or the second pressure surface G2.

[0086] The first heating block 310 and the second heating block 320 are arranged side by side, spaced apart from each other, and each has guide grooves 312 and 322 (see Figure 2) that pass through them. The first heating block 310 and the second heating block 320 are placed on guide members 331 and 332 that pass through 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, allowing the PCR plate 200 to be pressurized from above.

[0087] Of course, in this case, a constant temperature plate 350 is placed below the first heating block 310 and the second heating block 320, and regions having the same temperature as the first or second temperature provided by each heating block are made corresponding, so that the PCR plate can be pressurized from above and below.

[0088] As described above, the temperature control module 300 in the present invention has the advantage of being able to immediately apply the first temperature and the second temperature to the entire surface of the PCR plate 200, resulting in excellent effects in terms of application speed and reaction efficiency.

[0089] Furthermore, the heating block structure of the temperature control module 300 is positioned at the top where it can move horizontally at all times, and is designed to descend to the bottom only when it is pressed against the PCR plate 200.

[0090] To realize such a structure, the present invention may include a first elastic member S1 which is arranged as a structure inserted inside the drive frame and has a restoring force that keeps it constantly rising to the top when not being compressed. Furthermore, in the present invention, when pressurizing in contact with the PCR plate 200, a second elastic member 335 is provided to transmit the pressurizing force in order to prevent the application of excessive pressurizing force (Figure 5). In the illustrated embodiment, the second elastic member 335 is embodied as a leaf spring structure and exerts a certain buffering force when the first and second heating blocks 310 and 320 are pressed downwards, thereby controlling the application of excessive pressurizing force to the surface of the PCR plate 200.

[0091] Furthermore, in the present invention, the PCR plate 200 has the form of a plate-shaped structure in which reaction wells are realized on the upper surface, but in this case, the PCR plate 200 may be configured to further include a constant temperature plate 350 structure at the bottom so that it can be maintained at a certain temperature, for example, in the range of a second temperature (e.g., 55°C).

[0092] This is because, when the temperature is increased by the temperature control module 300 of the present invention while the first heating block at a first temperature (e.g., 95°C) is in close contact with the PCR plate 200, or when the second heating block at a second temperature (e.g., 55°C) is in close contact with the PCR plate 200, the internal amplification efficiency is significantly better only when the PCR plate 200 reaches the target temperature rapidly.

[0093] Therefore, embodiments of the present invention may further include a constant temperature plate 350 positioned below the PCR plate 200 to maintain the temperature of the PCR plate 200 at a second temperature.

[0094] In particular, the constant temperature plate 350 structure may be implemented by being fixed in place and applying a set temperature, but as described above, it can also be implemented by dividing the constant temperature plate 350 itself into areas for applying a first temperature and a second temperature, and by a structure that allows the constant temperature plate to be moved horizontally.

[0095] Figure 6 shows the structure in Figure 2, where the constant temperature plate 350 is moved horizontally through the horizontal movement drive module 400 to enter the lower part of the temperature control module. Figure 7 shows the operation in Figure 6 where the constant temperature plate 350 is moved horizontally outward, changing the temperature range.

[0096] Specifically, in the structure shown in Figure 6, when the first heating block 310 is positioned to apply a first temperature, the first region of the constant temperature plate 350 moves horizontally to the bottom of the PCR plate together with the first heating block, so that the first heating block 310 faces the top surface of the PCR plate 200. Subsequently, as shown in Figure 7, when the second region moves horizontally to the bottom of the PCR plate together with the second heating block, the second heating block 320 moves horizontally and operates in a manner that faces the top surface of the PCR plate 200.

[0097] The horizontal movement of the constant temperature plate 350 can be implemented using a sliding mechanism, and it can be implemented so that it moves while in contact with a sliding tape that is in contact with the side surface of the constant temperature plate 350.

[0098] Referring also to Figures 6 and 7, which are conceptual diagrams showing the bottom surface of the temperature control module 300 according to the present invention, the constant temperature plate 350 is positioned between the PCR plate 200 and the lower scanning module (not shown: reference numeral 500 in Figure 1), and multiple light-transmitting sections H may be provided in a through-structure to guide the light from the scanning module 500 so that excitation light irradiated from the scanning module is transmitted to the PCR plate 200 and fluorescence can be detected.

[0099] Therefore, in the present invention, the nucleic acid extraction, PCR process, and detection process can be implemented within a single system in an integrated system equipped with the scanner described above, and a separable PCR plate structure can be implemented to enable application to the diagnosis of various diseases.

[0100] Figures 8 to 13 show an example of a PCR plate 200 to which the present invention is applied.

[0101] Referring to Figure 8 and the conceptual diagrams in Figures 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 channel portion 230, and a blocking portion 240.

[0102] The torso section 210 is equipped with one or more reaction wells W (W1...Wn). The reaction wells W receive a PCR (polymerase chain reaction) preliminary mixture or nucleic acid solution (hereinafter referred to as "nucleic acid solution") from the nucleic acid extraction cartridge 100 on a plate-shaped surface and contain primers, primer / probes, or a dried PCR mixture containing primer / probes.

[0103] The insertion section 220 extends from the body section 210, is inserted into the nucleic acid extraction cartridge 100, and has an injection port h1 into which the nucleic acid solution is injected. The insertion section 220 is connected to the body section 210 by extending from one end and being inserted into the nucleic acid extraction cartridge 100.

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

[0105] In the present invention, the reaction well W is embodied in the structure shown in Figure 8, which comprises eight reaction wells W1 to W8. However, it is not limited to this, and it is naturally possible to embody a structure with one or more reaction wells. The structure of the reaction well W can also be embodied as a concave pattern structure by processing the surface of the body portion 210.

[0106] In particular, in the embodiments of the present invention, as shown in Figure 8, the surface area of ​​the torso portion 210 is provided with a structure that partitions a certain area of ​​reaction wells W, so that the primer, which is provided in a dry state in the reaction wells W, and the nucleic acid solution injected from the nucleic acid extraction cartridge 100 can be dispersed and mixed in the reaction wells W.

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

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

[0109] The blocking housing section 213 is formed in a concave shape in the body frame section 211 and houses the blocking section 240. The blocking housing section 213 is formed in a concave shape corresponding to the size of the blocking section 240 so that it can accommodate the blocking section 240.

[0110] The connecting channel section 215 connects the blocking containment section 213 to each reaction well W. The nucleic acid solution can be supplied to the reaction well W via the connecting channel section 215.

[0111] The storage section 217 is formed in a concave shape in the body section 210, communicates with the flow channel section 230, and receives the nucleic acid solution. The storage section 217 receives and stores the nucleic acid solution so that it can be supplied to each reaction well W.

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

[0113] The connecting channel section 215 includes a bottle neck section 215a that is formed to be narrower on the reaction well W side from the shut-off section 240. The bottle neck section 215a is formed to be abruptly narrower at the connecting channel section 215, and the portion in which the shut-off section 240 is housed in the shut-off housing section 213 protrudes slightly.

[0114] Referring to Figure 10, the shut-off section 240 is inserted into the shut-off housing section 213 while being pressurized from above to below, and the width of the bottle neck section 215a of the connecting flow path section 215 is formed to be narrow.

[0115] Referring to Figure 11, when the transparent film is sealed and attached to the neck portion 215a of the bottle by pressure, the structure is such that all four sides of the connecting channel portion 215 are pressurized by the blocking portion 240, thereby blocking the flow of the nucleic acid solution. Therefore, the flow of the nucleic acid solution is blocked by the weak pressure difference between each adjacent reaction well W.

[0116] Referring to Figure 12, when nucleic acid solution is continuously injected into the blocking section 240, it pushes the blocking section 240 as it passes from the storage section 217 through the flow path guide section 219, and the nucleic acid solution flows into each reaction well W.

[0117] Referring to Figure 13, after the nucleic acid solution flows into each reaction well W, the pressure rises due to the temperature change in each reaction well W, pressurizing the blocking section 240 in the reaction well W. The blocking section 240 then tightly seals and pressurizes the flow path guide section 219, thereby preventing the nucleic acid solution from flowing back from the reaction well W to the flow path section 230.

[0118] Furthermore, the PCR plate 200 is configured to include a cover member (not shown) that seals the top of a number of reaction wells W, and a transparent film material with light transmission properties can be applied to the cover member.

[0119] As the cover member adheres closely to the surface of the reaction well W and creates a cavity inside the reaction well W, the PCR (polymerase chain reaction) preliminary mixture and nucleic acid solution, which will later be injected from the nucleic acid extraction cartridge 100, are injected into the reaction well W while pushing out the air layer present in the cavity.

[0120] In particular, in the present invention, as shown in the structure of Figure 8, the flow channel section 230 connected to the reaction well W in the body section 210 is implemented to extend from the inlet h1 through the body section 210 to the end of the body section 210, and at the end, it is implemented to be connected to the ends of a number of reaction holes in the opposite direction to the insertion section 220.

[0121] In other words, as shown in Figure 2, when the PCR premixture and nucleic acid solution are injected into the nucleic acid extraction cartridge 100 via the injection port h1 provided at the bottom of the insertion section, the flow channel section 230 is realized in a direction x1 that crosses the body section 210, and can be realized so as to branch to the left and right from the end point of the body section 210 and connect to the inlet of each reaction well W. The reason for forming the flow channel in this way is that a small amount of air layer is present inside the reaction well W region sealed by the cover member, so that the injected PCR premixture and nucleic acid solution rise from the upper region with respect to the center line Cx of the body section as shown in the structure of Figure 8, and the air layer slides up to the lower region Cb of the body section 210.

[0122] Therefore, the mixture in which the PCR reaction is carried out is positioned relatively in the upper region Ca of the reaction well W. This means that, due to the characteristics of the equipment, the region in which the heating blocks 310 and 320 of the present invention apply pressure and the region in which the scanner module performs detection are located in the upper region Ca, as shown in Figure 8, thereby improving the precision of detection, the efficiency of the PCR reaction, and the efficiency of temperature control.

[0123] Furthermore, in the present invention, the PCR plate 200 can be made of a synthetic resin material with high light transmittance. This is to improve detection efficiency by using a material with high light transmittance due to the function of the scanner module described above.

[0124] While various synthetic resin materials such as transparent PP, PE, PPA, PMMA, and PC can be used for such materials, they are not necessarily limited to these; any material that can ensure a certain level of light transmittance can be used.

[0125] However, the PCR plate 200 is designed to maintain a constant temperature through a heat source applied from the lower constant temperature plate 350. To ensure efficient temperature maintenance of the temperature control module, which is applied directly to the PCR reaction product containing at least one of the PCR (polymerase chain reaction) premixture or nucleic acid solution, and dried primers and probes, the thickness of the body portion 210 is designed to be in the range of 1.0 mm to 3.0 mm. If the thickness of the body portion 210 is less than 1.0 mm, the high heat used to set the first temperature is easily transferred to the lower part of the body portion 210, causing thermal interference with the constant temperature plate 350 and making temperature control difficult. If the thickness of the body portion 210 exceeds 3.0 mm, temperature control of the substance contained in the reaction well W is easy, but temperature control of the lower constant temperature plate 350 becomes difficult, resulting in the disadvantage of difficulty in maintaining a constant temperature.

[0126] In other words, as described with reference to Figures 4 and 5, in the present invention, the first heating block 310 and the second heating block 320 are moved horizontally, and the first or second pressurizing surface G1 or G2 in contact with the surface of the reaction well W is pressurized in a structure that contacts the upper surface of the partition wall pattern and the cover member covering the partition wall pattern, thereby controlling the temperature of the reactants while setting the temperature to the first or second temperature.

[0127] Furthermore, when temperature circulation is performed on the PCR plate 200, in the temperature control module 300 with the structure shown in Figures 2 and 3, in order to raise the temperature to the first temperature, the first heating block 310 moves horizontally so that it faces the upper surface of the PCR plate 200, and on the lower surface of the PCR plate 200, the first region of the constant temperature plate 350 moves horizontally together with the first heating block 310, and then the first heating block 310 moves downward while making contact and applying pressure. In order to lower the temperature to the second temperature, the upper surface of the PCR plate 200 moves horizontally so that it faces the second heating block 320, and on the lower surface of the PCR plate 200, the second region of the constant temperature plate 350 moves horizontally together with the second heating block 320, and then the second heating block 320 moves downward while making contact and applying pressure, so that heating and cooling are performed simultaneously on the upper and lower surfaces of the PCR plate 200. In this invention, the constant temperature plate 350 and the first and second heating blocks 310 and 320 are implemented in a structure that moves horizontally together as a single unit, so it is natural that the upper and lower parts of the PCR plate 200 can be pressurized simultaneously at the same temperature.

[0128] This operation allows for simultaneous contact and pressurization of the top and bottom surfaces of the PCR plate 200, achieving twice the efficiency compared to the constant temperature plate 350 method, which maintains a single temperature. This method, which simultaneously heats the plate inside the target on both the top and bottom surfaces, offers the advantage of reducing testing time by half.

[0129] Figures 14 to 19 show other examples of the PCR plate 200 to which the present invention is applied.

[0130] Referring to Figure 14 and the conceptual diagrams in Figures 2 and 3 described above, the PCR plate 200 of another embodiment of the present invention includes a body portion 210, an insertion portion 220, a flow channel portion 230, and a blockage portion 240. The explanation of the body portion 210, insertion portion 220, flow channel portion 230, and blockage portion 240 of the PCR plate 200 is as described above.

[0131] In another embodiment of the PCR plate 200 of the present invention, the containment section 213 is divided into multiple sections by partitions 214 formed along the length of the storage section 217. The containment sections 240 are mounted in each of the containment sections 213 divided by the partitions 214.

[0132] Referring to Figure 16, multiple blocking units 240 are inserted into their respective blocking housings 213 from top to bottom. The elastically deformable blocking units 240 are press-fitted into the blocking housings 213. The elastically deformable blocking units 240 seal the blocking housings 213.

[0133] Referring to Figure 17, after being sealed by the blocking section 240, the flow due to the pressure difference in each blocking housing section 213 is blocked by the partition wall section 214.

[0134] Referring to Figure 18, when the nucleic acid solution is injected, it pushes the blocking section 240 as it passes through the storage section 217 and the connecting channel section 215, and flows into each reaction well W.

[0135] Referring to Figure 19, after the nucleic acid solution flows in, the pressure in each reaction well W increases due to the temperature change, pressurizing the blocking section 240 in the reaction well W. The blocking section 240 can block the backflow of the nucleic acid solution while tightly sealing and pressurizing the connecting channel section 215.

[0136] Figures 20 to 23 are diagrams illustrating in detail the structure and operating method of such a constant temperature freight and horizontal movement drive module.

[0137] Figure 20 shows the structure in which the constant temperature plate 350 is placed, as in Figures 2 and 3, while Figure 22 shows the structure with only the constant temperature plate structure separated.

[0138] Referring to Figures 20 and 21, the constant temperature plate 350 according to an embodiment of the present invention is positioned below the heating block 310 and 320 structures that constitute the temperature control module 300 shown in Figures 2 and 3. After the PCR plate 200 is placed, when the first heating block 310 and the second heating block 320 of the temperature control module 300 pressurize the PCR plate 200 from above by horizontal and vertical movement, the constant temperature plate 350 functions to maintain the same temperature as the first heating block 310 and the second heating block 320.

[0139] As shown in Figure 20, the constant temperature plate 350 is provided with a separation section Ss that divides the first region a1, which maintains a first temperature, and the second region a2, which maintains a second temperature, and the first region a1 and the second region a2 are connected with respect to the two side ends a3 and a4 of the separation section Ss.

[0140] A connector structure Ca and Cb is attached to one end of the constant temperature plate 350, allowing power to be applied or control signals to be transmitted.

[0141] In particular, the horizontal movement of the constant temperature plate 350 is implemented using a sliding mechanism, and it moves by contacting a sliding tape that contacts the side surface of the constant temperature plate 350. This not only simplifies the structure but also enhances mobility.

[0142] In this case, the second region a2 can also be equipped with a light-transmitting section H so that the detection light from the scanning module 500, which scans the concentration of the amplified reactants, can be transmitted through it.

[0143] The first region a1 and the second region a2 are equipped with temperature sensors Sa and Sb, respectively, allowing for the measurement and control of the temperature in the corresponding regions.

[0144] Figure 23 shows the lower view of Figure 22, and is provided with connection connectors Cc and Cd for applying control signals and power supply, and is equipped with temperature sensors Sa and Sb to enable constant temperature maintenance of the first and second temperatures.

[0145] To maintain the temperature of the first and second regions of the constant temperature plate 350, various heating means, such as heating wires and heating antibodies, can be installed inside the plate. However, in a preferred embodiment of the present invention, circuits for electrodes and temperature sensors are realized in an epoxy printed circuit, and after applying a heat-generating paint between each electrode, metal plates corresponding to the first and second regions are bonded so as to be in close contact with their respective temperature sensors and heat-generating paint, thereby realizing such an effect.

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

[0147] As described above, the horizontal movement drive module 400 can be configured to include a moving bar 420 and a drive motor unit 410 connected to one end of the constant temperature plate 350, as shown in Figures 2 and 3, 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.

[0148] As shown in Figure 21, the PCR plate 200 is inserted into the nucleic acid extraction cartridge 100 and connected in a structure that connects the channels, and the nucleic acid solution extracted from the nucleic acid extraction cartridge 100 is injected into the injection port h1, and the injected extracted nucleic acid solution then moves into the PCR plate 200 which contains one or more reaction wells containing a PCR mixture dry product containing at least one side of the primer and probe.

[0149] Subsequently, the first heating block 310 and the second heating block 320 of the temperature control module 300 of the present invention, which are used to establish a first or second temperature in the reaction well portion of the PCR plate 200, begin to descend.

[0150] In this case, the horizontal movement drive module 400 moves both the constant temperature plate 350 and the temperature control module 300 horizontally, and the PCR plate 200 is positioned to be inserted between the constant temperature plate 350 and the temperature control module 300.

[0151] When the heating blocks 310 and 320, which have been moved horizontally via the horizontal movement drive module 400, pressurize the PCR plate 200, the first or second region of the constant temperature plate 350, which has a set temperature (first or second temperature) corresponding to the temperature of the heating block 310 or 320, is positioned to naturally correspond to it.

[0152] In other words, when the first region a1 of the constant temperature plate 350 moves horizontally to the bottom of the PCR plate 200, the first heating block (Figure 2) 310 moves horizontally at the same time, so as to face the top surface of the PCR plate 200, the heating blocks 310 and 320 then descend and come into contact with the top surface of the PCR plate 200.

[0153] Furthermore, when the second region a2 moves horizontally to the bottom of the PCR plate 200, the second heating block (Figure 2) 320 simultaneously moves horizontally and operates in a corresponding manner to face the top surface of the PCR plate 200, after which the heating blocks 310 and 320 descend and come into contact with the top surface of the PCR plate 200.

[0154] In short, when performing temperature circulation on the PCR plate 200, in order to raise it to the first temperature, the first heating block 310 moves horizontally so that it faces the upper surface of the PCR plate 200, and the lower surface of the PCR plate 200 is driven so that it comes into contact with and is pressurized by the first heating block 310 as it moves downward after the first region of the constant temperature plate 350 has moved horizontally.

[0155] Furthermore, in order to lower the temperature back to the second temperature, the upper surface of the PCR plate 200 moves horizontally so that it faces the second heating block 320, and on the lower surface of the PCR plate 200, the second region of the constant temperature plate 350 moves horizontally together with the second heating block 320, after which the second heating block 320 moves downward and is brought into contact with and pressurized, so that heating and cooling occur simultaneously on the upper and lower surfaces of the PCR plate 200.

[0156] In this way, by simultaneously applying contact and pressure to the upper and lower surfaces of the PCR plate 200, twice the efficiency is achieved compared to the constant temperature plate 350 method, which maintains a single temperature.

[0157] In the following, a nucleic acid extraction cartridge 100, which embodies a PCR (polymerase chain reaction) preliminary mixture or nucleic acid solution containing a nucleic acid extract in the PCR plate 200 of the present invention described above, will be explained with reference to Figures 24 to 32.

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

[0159] Referring to Figures 24 to 26, the nucleic acid extraction cartridge 100 according to the present invention may be configured to include a cartridge cover R1 having a number of partitioned containment sections 22, 23, 24, 25, 26 containing a solution necessary for DNA extraction, and a cartridge body R2 which is connected to the lid R1 by an insertion structure and is provided with a reaction containment section 11 for reacting or purifying the solution drawn in from each containment section 22, 23, 24, 25, 26 with a sample.

[0160] In this case, the system is configured to include a piston 18 for injecting the PCR preliminary mixture or nucleic acid solution purified in the reaction containment section 11 into the injection port h1 of the PCR plate 200, which is attached to the cartridge body section R2 in a structure that is inserted into the cartridge body section R2.

[0161] In the present invention, the cartridge cover R1 contacts one side surface of each reaction housing portion 11 of the cartridge body R2 and includes a rubber portion 30 made of an elastically deformable material. The rubber portion 30 is made of an elastically deformable material and includes rubber, silicone, and the like.

[0162] In the present invention, the rubber portion 30, when assembling the cartridge cover R1 and the cartridge body R2, seals the space between the cartridge cover R1 and the cartridge body R2, thereby preventing the nucleic acid solutions from mixing with each other by allowing the rubber portion 30 to ride up onto the wall surface of the reaction containment portion 11 of the cartridge body R2.

[0163] The operation of the nucleic acid extraction cartridge of the present invention will be explained below with reference to Figures 25 to 32. Figure 27 is a transparent perspective view of Figure 24, showing the internal structure after binding.

[0164] The nucleic acid extraction cartridge of the present invention is equipped with a rotary valve 19 located on the bottom surface of the main body R2, with a flow path 19-1 formed inside. By rotating this rotary valve, the spaces of each of the storage compartments 11, 12, 13, 14, 15, 16, and 17 of the cartridge main body R2 can be connected to the flow path of the rotary valve 19. After connecting the flow path to a specific storage compartment, the design allows the solution that has entered the storage compartment to be collected by operating the piston 18 and the substance to be transferred to another storage compartment or a PCR plate 200.

[0165] As shown in Figures 25 and 26, the cartridge cover R1 has internal storage compartments 22, 23, 24, 25, and 26 containing the respective solutions necessary for DNA extraction. The bottom of these storage compartments is sealed with a film or the like, so they are designed to be easily pierced by the needle of the main storage compartment. In addition, five holes 21-1, 21-2, 27, 28, and 29 are formed inside.

[0166] The first storage section 22 of the cartridge cover R1 contains the binding buffer, the second storage section 23 contains the first washing buffer, the third storage section 24 contains the second washing buffer, the fourth storage section 25 contains the third washing buffer, and the fifth storage section 26 contains the elution buffer.

[0167] PCR plate 200 is covered with a transparent plastic film (polyethylene, polypropylene, PET, etc.), and the reaction well contains a PCR dry product mixture that includes at least one side of the dried PCR primers and probes. This structure is the same as the one described with reference to Figure 8.

[0168] The nucleic acid extraction cartridge of the present invention may operate in the following sequence.

[0169] 1. Addition of biological sample

[0170] The cartridge body R2, cartridge cover R1, and PCR plate 200 are assembled and mounted in the automated equipment described later, and a biological sample (blood) is placed into the first well 21-1 shown in Figure 25.

[0171] 2. Nucleic acid leakage from cells and binding to beads

[0172] As shown in Figure 30, the binding buffer in the first containment section 22 is introduced into the reaction containment section 11 through the rotation of the rotary valve 19 located at the bottom of the cartridge body section R2 and the action of the piston 18, and is mixed with the biological sample and the bead of the magnetic tablet MT (silica-coated magnetic bead).

[0173] The magnetic tablet MT used in this invention is attached to the end of a through-pipe that extends into the reaction containment section 11 of the cartridge body R2, and functions to dissolve nucleic acids extracted from cells contained in the biological sample onto the magnetic tablet and bind to the surface of the dispersed magnetic beads.

[0174] In this case, instead of the magnetic tablet, the magnetic bead may be suspended in the binding buffer and used.

[0175] Subsequently, when ultrasonic waves are applied while a sonication tip is inserted into the sealed second hole 21-2 of the cartridge body R2, the ultrasonic waves are transmitted through the plastic, and the reaction solution is homogenized as the biological sample, tablet, and binding buffer are mixed. At this time, the biological tissues contained in the biological sample are also broken down, causing nucleic acids to leak out, and these leaked nucleic acids bind to the surface of the bead.

[0176] When the magnetic bar is inserted into the third hole 27 of the cartridge body R2, the bead is fixed to the wall of the reaction containment section, and the remaining reaction liquid is transferred to the first containment section through the rotation of the rotary valve and the action of the piston.

[0177] 3.1 Primary Cleaning

[0178] The primary washing buffer from the second containment section 23 is introduced into the reaction containment section 11 through the rotation of the rotary valve and the action of the piston in the cartridge body section R2 shown in Figure 27, and mixed with the nucleic acid-bound beads.

[0179] Subsequently, the magnetic bar is removed from the third hole 27 in Figure 25, and a transducer is inserted into the second hole 21-2 while ultrasound is applied, performing a primary wash. This primary wash removes not only nucleic acids but also other substances that are nonspecifically bound to the bead.

[0180] As the magnetic bar is inserted into the third hole 27, the bead is fixed to the wall of the reaction containment section, and the primary cleaning solution is transferred to the second containment section 23 through the rotation of the rotary valve and the action of the piston.

[0181] 4.2 Secondary cleaning

[0182] As shown in Figure 27, the secondary washing buffer from the third containment section 24 is introduced into the reaction containment section 11 through the rotation of the rotary valve and the action of the piston in the cartridge body section R2, and mixed with the nucleic acid-bound beads.

[0183] Subsequently, the magnetic bar is removed from the third hole 27 in Figure 25, and a transducer is inserted into the second hole 21-2 while ultrasound is applied, performing a secondary cleaning. This secondary cleaning removes not only nucleic acids but also other substances that are nonspecifically bound to the bead.

[0184] As the magnetic bar is inserted into the third hole 27, the bead is fixed to the wall of the reaction containment section, and the secondary cleaning fluid is transferred to the third containment section 24 through the rotation of the rotary valve and the action of the piston.

[0185] 5.3 Secondary Cleaning

[0186] The tertiary washing buffer from the fourth containment section 25 is introduced into the reaction containment section 11 through the rotation of the rotary valve and the action of the piston in the cartridge body section R2 shown in Figure 27, and mixed with the nucleic acid-bound beads.

[0187] Subsequently, the magnetic bar is removed from the third hole 27 in Figure 25, and a transducer is inserted into the second hole 21-2 while ultrasound is applied, performing a tertiary cleaning. This tertiary cleaning removes not only nucleic acids but also other substances that are nonspecifically bound to the bead.

[0188] As the magnetic bar is inserted into the third hole 27, the bead is fixed to the wall of the reaction containment section, and the tertiary cleaning fluid is transferred to the fourth containment section 25 through the rotation of the rotary valve and the action of the piston.

[0189] 6. Nucleic acid elution

[0190] The elution buffer in the fifth containment section 26 is introduced into the reaction containment section 11 through the rotation of the rotary valve and the action of the piston in the cartridge body section R2 shown in Figure 27, and mixed with the nucleic acid-bound beads.

[0191] Subsequently, the magnetic bar is removed from the third hole 27 in Figure 25, and when ultrasound is applied while a transducer is inserted into the second hole 21-2, the nucleic acids bound to the surface of the bead are dissolved in the elution buffer.

[0192] 7. Preparation of PCR Premixture

[0193] As a magnetic bar is inserted into the third well 27, the bead is fixed to the wall of the reaction containment section. Through the rotation of the rotary valve and the selection of a small piston, the elution buffer containing dissolved nucleic acids is introduced into the sixth containment section 17 and mixed with the PCR material [a mixture of polymerase, dNTPs, etc.] that has entered the sixth containment section, thereby generating a PCR premixture. The "PCR premixture" used in this invention is defined and used as being embodied in the above-mentioned substances.

[0194] 8. Transfer to PCR plate

[0195] The PCR premixture and nucleic acid solution generated in the sixth containment section are introduced into the PCR plate 200 through the rotation of the rotary valve and the action of the piston in the cartridge body R2 shown in Figure 27, and are mixed with at least one of the primers and probes in the PCR plate 200. This introduction process is carried out by the pressurization of the piston 18, which moves along the flow path Y of the rotary valve and is injected into the injection port h1, as shown in Figure 32.

[0196] Subsequently, a heating rod is inserted into the fourth well 29, while the cover film at the entrance of the PCR reaction plate is pressurized and heated, thereby sealing the PCR reaction plate.

[0197] 9. PCR reaction

[0198] Ultimately, PCR plate 200 will contain nucleic acids extracted from the biological sample, polymerase, dNTPs, primers, probes (at least one of these), and other buffers.

[0199] Therefore, the PCR reaction is carried out by applying pressurized heat to the PCR plate 200 through the temperature control module of the present invention described above.

[0200] Figures 33 to 36 show the overall structure and arrangement of the apparatus constituting the polymerization enzyme chain reaction system of the present invention described above.

[0201] The polymerization enzyme chain reaction apparatus according to the embodiment of the present invention may include a nucleic acid extraction cartridge assembly comprising a nucleic acid extraction cartridge 100 and a PCR plate 200.

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

[0203] Figure 34 is an enlarged view of the combined arrangement diagram of the main parts of the present invention shown in Figure 33, and Figure 35 is a conceptual vertical cross-sectional view of the part shown in Figure 34, illustrating the arrangement of the main components. Figure 36 is a conceptual side perspective cross-sectional view of Figure 35.

[0204] As shown in Figures 34 to 36, the PCR preliminary mixture or nucleic acid solution containing nucleic acids extracted from inside the nucleic acid extraction cartridge 100 according to the present invention is injected into a PCR plate 200 on which reaction wells W are formed. A first pressurized surface G1 corresponding to the surface of the reaction wells W is formed on the upper part of the PCR plate 200, and a first heating block 310, which is maintained at the temperature required for thermal denaturation by a heating unit, is positioned there, and a second heating block 320, which can be formed in a structure in which the area being pressurized changes as it moves horizontally, is positioned nearby. As a result, the PCR preliminary mixture and nucleic acid solution injected into the reaction wells W are directly heated by the heating blocks to a first temperature (95°C) required for thermal denaturation and a second temperature (55°C) required for annealing.

[0205] In addition, as shown in Figures 35 and 36, a constant temperature plate 350 is placed below the PCR plate 200 to maintain the temperature of the PCR plate 200 at a constant temperature level.

[0206] A scanning module 500 is positioned at the bottom of the constant temperature plate 350, and light L irradiated by the light irradiation unit E1 passes through the light-transmitting unit H of the constant temperature plate 350 to reach the PCR plate 200, where fluorescence detection is performed.

[0207] In the embodiments of the present invention, as described above, when the temperature is increased while the first heating block 310 at a first temperature (e.g., 95°C) is in close contact with the temperature control module 300, or when the second heating block 320 at a second temperature (e.g., 55°C) is in close contact with the temperature control module 300, temperature control of the internally amplified reactants becomes much easier if the PCR plate 200 is maintained within a constant temperature range. Therefore, maintaining the temperature of the constant temperature plate 350 at the second temperature becomes a very important factor in increasing the reliability of the reaction. When raising the temperature of the PCR plate 200 to 95°C, the heat transfer rate is increased while the first temperature block and the first constant temperature area are in close contact, allowing the temperature of the PCR plate to reach 95°C rapidly within 2 to 3 seconds.

[0208] When performing RT / PCR to detect RNA targets, use a PCR premix or PCR plate containing a dried RT-PCR reaction product. After adjusting the low-temperature block to the RT reaction temperature and placing it in close contact with the PCR reaction plate, maintain it for the RT reaction time, perform the reverse transcription, and then perform the PCR reaction.

[0209] The presence or absence of amplified nucleic acid or its concentration can be determined through the PCR reaction, and this information can be used for diagnosis. In this case, the presence or absence or concentration of amplified nucleic acid can be determined using a standard nucleic acid detection method.

[0210] For example, methods using SYBR green, a DNA minor groove insertion fluorescent dye that performs DNA insertion, or methods using probes with various phosphors and quenchers attached to scan excitation light of various wavelengths with the corresponding fluorescence are available, and are not limited to these.

[0211] Referring to Figures 37 to 44, in yet another embodiment of the present invention, the body portion 210 of the PCR plate 200 includes a body frame portion 211, a containment portion 213, and a connecting channel portion 215. The body frame portion 211 comprises one or more reaction wells W (W1...Wn).

[0212] The blocking housing section 213 is formed in a concave shape in the body frame section 211, is positioned corresponding to each reaction well W, and is circular in shape to accommodate the circular blocking section 240. The blocking housing section 213 is formed in a concave shape corresponding to the size of the blocking section 240 so that it can accommodate the blocking section 240.

[0213] The connecting channel section 215 connects the blocking containment section 213 to each reaction well W. The nucleic acid solution can be supplied to the reaction well W via the connecting channel section 215.

[0214] The blocking section 240 is made of an elastically deformable material. The blocking section 240 is made of an elastically deformable material such as rubber or silicone. When the nucleic acid solution is injected into each reaction well W, the blocking section 240 placed in each blocking housing section 213 is made of rubber or the like and has little weight, so the resistance applied to each blocking housing section 213 is small.

[0215] Therefore, the nucleic acid solution is injected through a single channel 230, and the same amount of nucleic acid solution is uniformly injected into each reaction well W.

[0216] Referring to Figures 43 and 44, a hole is formed below each blocking housing 213, and the blocking unit 240 contacts this hole by its own weight. When the pressure in each reaction well W becomes higher than that on the connecting flow channel 215 side, the blocking unit 240 tightly seals against the hole in the blocking housing 213, and can function as a check valve to prevent backflow of nucleic acid solution.

[0217] Figure 45 shows the results of testing in reaction well W, specifically in the outermost reaction wells W1 and W8 out of the eight reaction wells W1 to W8. To confirm whether the nucleic acid solutions were mixed between the reaction wells W during the PCR reaction, the reaction wells were dried before proceeding with the PCR.

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

[0219] Figure 46 shows the results of testing in reaction well W, specifically in the outermost reaction wells W1 and W8 out of the eight reaction wells W1 to W8. After drying the outermost reaction wells, PCR was performed, and it was confirmed that the check valve prevented the solutions between the reaction wells from mixing.

[0220] Referring to Figures 37 to 41, the nucleic acid extraction cartridge 100 has holes 21-1, 21-2, 27, 28, and 29. Leak-proof rubber 40 is fitted to each of the holes 21-1, 21-2, 27, 28, and 29 of the nucleic acid extraction cartridge 100. The leak-proof rubber 40 is made of an elastic material such as rubber.

[0221] This leak-proof rubber 40 prevents residual nucleic acid solution in the nucleic acid extraction cartridge 100 from leaking through the holes 21-1, 21-2, 27, 28, and 29 of the nucleic acid extraction cartridge 100.

[0222] Referring to Figure 39, the leak-proof rubber 40 is punched by the crescent-shaped punch 50, creating a hole through which air can enter.

[0223] Referring to Figure 40, when the nucleic acid solution flows in, the leak-proof rubber 40 is pressed by the pressing jig 60, and atmospheric pressure can be smoothly drawn in through the holes in the leak-proof rubber 40.

[0224] Referring to Figure 41, when the nucleic acid extraction cartridge 100 is disposed of, the leak-proof rubber 40 can close the nucleic acid extraction cartridge 100 and prevent leakage of the nucleic acid solution remaining inside the nucleic acid extraction cartridge 100.

[0225] Referring to Figure 44, the sheet portion 250 is heat-sealed to the reaction well W. By heat-sealing each reaction well W with the sheet portion 250, the volume of each reaction well W is maximized, allowing heat transfer within the reaction well W to proceed rapidly, and enabling the reaction of the nucleic acid solution to proceed quickly.

[0226] Although the present invention has been described with reference to one embodiment shown in the drawings, this is merely illustrative, and a person with ordinary skill in the art will understand that a variety of modifications and equivalent other embodiments are possible. Therefore, the true scope of technical protection of the present invention should be defined by the following claims.

Claims

1. A torso having one or more reaction wells; An insertion portion extending from the aforementioned body portion, which is inserted into a nucleic acid extraction cartridge and has an injection port into which a nucleic acid solution is injected; A flow channel that allows the nucleic acid solution to flow from the inlet into the reaction well; and A blocking part attached to the body portion is included to block the backflow of the nucleic acid solution from the reaction well to the flow channel portion; The aforementioned torso section is, A fuselage frame portion comprising one or more of the aforementioned reaction wells; A storage section formed in a concave shape in the body frame, communicating with the flow channel section, and receiving the supply of the nucleic acid solution; A shielding housing portion formed in a concave shape in the body frame portion, which houses the shielding portion; A flow path guide section that communicates with the storage section and guides the nucleic acid solution to flow toward the blocking containment section; and A connecting channel section connecting the blocking containment section and the reaction well; When the pressure in the reaction well rises, the blocking section is pressurized and adheres tightly to the flow path guide section, preventing the backflow of nucleic acid solution from the reaction well to the flow path guide section. A PCR plate characterized by the following features.

2. The PCR plate according to claim 1, characterized in that the connecting channel portion includes a bottle neck portion that is formed to be narrower in width from the blocking portion toward the reaction well.

3. The aforementioned containment section is divided into multiple partition sections formed along the length of the storage section. The PCR plate according to claim 1, characterized in that the blocking portion is attached to each of the divided blocking housing portions.

4. The PCR plate according to claim 1, characterized in that the barrier portion is made of an elastically deformable material.

5. A fuselage section equipped with numerous reaction wells; An insertion portion extending from the aforementioned body portion, which is inserted into a nucleic acid extraction cartridge and has an injection port into which a nucleic acid solution is injected; A flow channel that allows the nucleic acid solution to flow from the inlet into the reaction well; and A blocking part attached to the body portion is included to block the backflow of the nucleic acid solution from the reaction well to the flow channel portion; The aforementioned torso section is, A fuselage frame portion comprising one or more of the aforementioned reaction wells; A concave section formed in the body frame, arranged corresponding to each of the reaction wells, and formed circularly to accommodate the circular section; and A connecting channel that connects the blocking containment section and the reaction well; Includes, The blocking portion includes an elastically deformable material, and when the pressure in the reaction well rises above the pressure in the connecting channel portion, the blocking portion adheres tightly to the hole in the blocking housing portion to prevent backflow of the nucleic acid solution. A PCR plate characterized by the following features.

6. The PCR plate according to claim 5, characterized in that a sheet portion is heat-sealed to the reaction well.

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