PCR reaction apparatus and reagent detection system

Abstract

This application relates to a PCR reaction apparatus and a reagent detection system. The PCR reaction apparatus of this application includes: a frame, a reagent kit, a first temperature control mechanism, a second temperature control mechanism, and a motion mechanism. The reagent kit is disposed on the frame and has at least one functional well. The first temperature control mechanism is disposed on the frame and / or the reagent kit. The second temperature control mechanism is disposed above the reagent kit and includes a temperature adjusting element and a temperature conducting element. The temperature adjusting element is used to adjust the temperature of the temperature conducting element. The motion mechanism is disposed on the frame and is drivenly connected to the temperature conducting element to drive the temperature conducting element closer to or away from the functional well. This application can adjust the temperature of the items stored in the functional well of the reagent kit through the first temperature control mechanism and the second temperature control mechanism, respectively. The temperature adjustment ranges of the first temperature control mechanism and the second temperature control mechanism are different to meet the various needs of general nucleic acid detection and methylation.

Description

Technical Field

[0001] This application relates to the technical field of gene detection, and more specifically, to a PCR reaction apparatus and reagent detection system. Background Technology

[0002] Epigenetics encompasses numerous types of epigenetic modifications, including DNA methylation, genomic imprinting, maternal effects, gene silencing, nucleolar dominance, activation of dormant transposons, and RNA editing. DNA methylation is the most widely used. It refers to the process by which an organism, under the catalysis of DNA methyltransferases, transfers a methyl group to a specific base using S-adenosylmethionine as a methyl donor. Bisulfite sequencing is currently the most commonly used and accurate method, capable of performing whole-genome methylation sequencing and DNA methylation detection, meeting various needs for tumor risk prediction. Furthermore, it has been applied in plant science, demonstrating significant effectiveness in screening and breeding.

[0003] As methylation becomes more widely used, more and more automated nucleic acid detection reagent systems are becoming compatible with methylation detection. Methylation often requires multiple extractions of the sample, and there are complex temperature control requirements during the extraction process. The difficulty and complexity of manually completing such operations is too great. Therefore, full automation of related reagent detection systems is an inevitable trend, especially in the transfer and pretreatment of nucleic acid products. Having the reagent detection system complete some key operations can greatly reduce the risk of contamination caused by manual operation.

[0004] However, existing PCR reaction devices generally only have one temperature control mechanism, which can only meet the needs of nucleic acid detection. When methylation is required, it is often necessary to transfer the PCR tube to a dedicated temperature control device to achieve methylation, which is inefficient. Furthermore, the frequent opening and closing of the PCR tube and the transfer of the tubes will increase the risk of sample contamination. Summary of the Invention

[0005] The purpose of this application is to provide a PCR reaction apparatus and reagent detection system, which can adjust the temperature of test tubes through a first temperature control mechanism and a second temperature control mechanism respectively.

[0006] The embodiments of this application are implemented as follows:

[0007] In a first aspect, this application provides a PCR reaction apparatus, comprising: a frame, a reagent kit, a first temperature control mechanism, a second temperature control mechanism, and a motion mechanism. The reagent kit is disposed on the frame and has at least one functional well. The first temperature control mechanism is disposed on the frame and / or the reagent kit. The second temperature control mechanism is disposed above the reagent kit and includes a temperature adjusting element and a temperature guiding element. The temperature adjusting element is used to adjust the temperature of the temperature guiding element. The motion mechanism is disposed on the frame and is connected to the temperature guiding element for driving the temperature guiding element to move closer to or away from the functional well.

[0008] In one embodiment, the PCR reaction apparatus further includes a clamping element configured to be close to or away from the functional well.

[0009] In one embodiment, the PCR reaction apparatus further includes a clamping member drive mechanism, which is disposed on the frame and connected to the clamping member for driving the clamping member to move.

[0010] In one embodiment, the clamping member includes: a main plate configured to be pushed by a temperature-conducting element; and at least one pressure rod connected to the main plate. Multiple functional holes are provided for placing test tubes, each test tube comprising a row of interconnected PCR tubes. Each PCR tube has a groove on its outer wall, and a connecting rod is provided between adjacent PCR tubes. The pressure rod is configured to extend into the groove and press against the connecting rod.

[0011] In one embodiment, the main body plate is provided with a drive groove into which the temperature-conducting element extends.

[0012] In one embodiment, the PCR reaction apparatus further includes: a first slide rail and a first slider, the first slide rail being disposed on the frame and located on one side of the reagent kit; the first slider being slidably disposed on the first slide rail and connected to the main body plate.

[0013] In one embodiment, a plurality of functional holes are provided, including at least one cap storage hole for placing a test tube cap and at least one tube storage hole for placing a test tube; wherein, the temperature-conducting element is configured to connect with the test tube cap to drive the test tube cap to move.

[0014] In one embodiment, the PCR reaction apparatus further includes a cap removal mechanism connected to the motion mechanism for separating the temperature-conducting element from the test tube cap.

[0015] In one embodiment, the motion mechanism includes: a translation mechanism, a lifting mechanism, and a mounting base, with the mounting base located above the frame; the lifting mechanism is located on the mounting base and is connected to the temperature-conducting component for driving the temperature-conducting component to rise and fall; the translation mechanism is located on the frame and is connected to the mounting base for driving the mounting base to translate; wherein, the cover-removing mechanism includes at least one cover-removing plate, which is fixed to the mounting base and arranged along the axis of the temperature-conducting component.

[0016] Secondly, this application provides a reagent detection system, including: a chassis, a robotic arm device, and a PCR reaction device as described in any of the foregoing embodiments, wherein the robotic arm device is disposed inside the chassis; and the PCR reaction device is disposed inside the chassis.

[0017] The advantages of this application over the prior art are:

[0018] The PCR reaction apparatus 17 of this application can adjust the temperature of the items stored in the functional wells of the reagent kit through a first temperature control mechanism and a second temperature control mechanism, respectively. The first and second temperature control mechanisms have different temperature ranges to meet various needs of routine nucleic acid detection and methylation. When applied to a reagent detection system, it eliminates the need to transfer PCR tubes and other items to a dedicated temperature control device for methylation when required, improving the efficiency of the reagent detection system and avoiding the risk of sample contamination caused by frequent opening, closing, and transferring of PCR tubes and other items.

[0019] Furthermore, this application incorporates a clamping component that can clamp the test tubes, preventing the test tubes or caps from leaving the reagent kit under the drive of the temperature-conducting component.

[0020] In addition, the temperature-conducting component of the second temperature control mechanism in this application can be connected to the test tube cap to drive the test tube cap to move, realize the automatic opening and closing of the PCR tube cap, and transfer the PCR tube cap to achieve automation, which can reduce manual intervention and eliminate the need for a separate opening and closing mechanism for the PCR tube in the reagent detection system. This simplifies the overall structure of the reagent detection system, makes the structure more compact, and reduces costs. Attached Figure Description

[0021] To more clearly illustrate the technical solutions of the embodiments of this application, the accompanying drawings used in the embodiments will be briefly introduced below. It should be understood that the following drawings only show some embodiments of this application and should not be regarded as a limitation of the scope. For those skilled in the art, other related drawings can be obtained based on these drawings without creative effort.

[0022] Figure 1 This is a top view of a reagent detection system shown in one embodiment of this application.

[0023] Figure 2 This is a schematic diagram of the internal structure of a reagent detection system according to an embodiment of this application.

[0024] Figure 3 This is a schematic diagram of the structure of a PCR reaction apparatus shown in one embodiment of this application.

[0025] Figure 4 This is a schematic diagram of an explosion of a PCR reaction apparatus according to an embodiment of this application.

[0026] Figure 5 This is a schematic diagram of an explosion of a PCR reaction apparatus according to an embodiment of this application.

[0027] Figure 6 This is a front view of a PCR reaction apparatus shown in one embodiment of this application.

[0028] Icons: 10-Reagent testing system; 11-Chassis; 12-Robotic arm device; 121-First robotic arm; 122-Second robotic arm; 123-Third robotic arm; 13-Centrifuge device; 14-PCR testing device; 15-Nucleic acid extraction device; 16-Sample processing device; 17-PCR reaction device; 200-Rack; 300-Reagent kit; 310-Functional wells; 311-Storage cap hole; 312-Storage tube hole; 301-PCR tube area; 302-Reagent tube area; 303-Sample retention tube area; 400-First temperature control mechanism; 500-Second temperature control mechanism; 510-Temperature regulator; 520-Temperature conductor; 521-Card slot ; 530-Temperature sensor; 540-Overheat switch; 600-Motion mechanism; 610-Mounting base; 620-Lifting mechanism; 621-Lifting motor; 622-Belt pulley transmission mechanism; 630-Translation mechanism; 640-Second slide rail; 650-Second slider; 700-Clamping component; 710-Main plate; 711-Drive groove; 720-Pressure rod; 730-Clamping component drive mechanism; 740-First slide rail; 750-First slider; 800-Cap removal mechanism; 801-Cap removal plate; 910-PCR tube; 911-Groove; 912-Connecting rod; 920-PCR tube cap; 921-Connecting groove; 922-Spring. Detailed Implementation

[0029] The terms “first,” “second,” “third,” etc., are used only for distinguishing descriptions and do not indicate a sequence number, nor should they be interpreted as indicating or implying relative importance.

[0030] Furthermore, terms such as "horizontal," "vertical," and "sag" do not imply that components must be absolutely horizontal or suspended, but rather that they can be slightly tilted. For example, "horizontal" simply means that its direction is more horizontal relative to "vertical," and does not mean that the structure must be completely horizontal, but can be slightly tilted.

[0031] In the description of this application, it should be noted that the terms "inner", "outer", "left", "right", "upper", "lower", etc., indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings, or the orientation or positional relationship commonly used when the product of this application is in use. They are only for the convenience of describing this application and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation. Therefore, they should not be construed as limitations on this application.

[0032] In the description of this application, unless otherwise expressly specified and limited, the terms “set up,” “install,” “connect,” and “link” shall be interpreted broadly, for example, as a fixed connection, a detachable connection, or an integral connection; as a mechanical connection or an electrical connection; as a direct connection or an indirect connection through an intermediate medium; or as a connection within two components.

[0033] Genetic testing is a medical testing technology that extracts nucleic acids from the peripheral venous blood, tissues, and other bodily fluids of the person being tested. The testing equipment then analyzes the DNA or RNA molecules in the person's cells to understand their genetic information and thus determine the cause or risk of disease.

[0034] Nucleic acid extraction is a pretreatment process for gene detection. Based on the known nucleic acid sequence, specific primers and probes are designed and synthesized. The extracted nucleic acid is then used as a template for quantitative real-time PCR (qPCR) experiments. The fluorescence signal is used to determine the positive or negative result of the target sample. Nucleic acid extraction is a crucial step in gene detection, and the quality of the obtained nucleic acid directly affects the success or failure of downstream experiments.

[0035] The technical solution of this application will now be clearly and completely described with reference to the accompanying drawings.

[0036] Please refer to Figure 1 This is a top view of the reagent detection system 10 shown in one embodiment of this application. Please refer to... Figure 2 This is a schematic diagram of the internal structure of a reagent detection system 10 according to an embodiment of this application. A reagent detection system 10 includes: a chassis 11, and the chassis 11 is provided with a robotic arm device 12, a centrifuge device 13, two PCR detection devices 14, a nucleic acid extraction device 15, a sample processing device 16, and a PCR reaction device 17.

[0037] The robotic arm device 12 may include three robotic arms: a first robotic arm 121, a second robotic arm 122, and a third robotic arm 123. The first robotic arm 121 is equipped with a cover opening and closing assembly. The sample processing device 16 includes a sample rack and a cup clamping mechanism. The first robotic arm 121 is used to move the sample tubes stored on the sample rack to the cup clamping mechanism for sample dispensing and other operations. The second robotic arm 122 is equipped with a pipette for pipetting. The third robotic arm 123 is equipped with an electric gripper or a moving part that can be connected to the test tubes for transferring test tubes such as PCR tubes 910 between the PCR reaction device 17, the nucleic acid extraction device 15, the centrifugation device 13, and the PCR detection device 14 to achieve functions such as PCR reaction, nucleic acid extraction, centrifugation, or PCR detection.

[0038] PCR reaction apparatus 17 includes: rack 200, first temperature control mechanism 400, and second temperature control mechanism 500 (please refer to...). Figure 3 The frame 200 is fixed inside the chassis 11 by bolts or other means. The frame 200 houses the reagent kit 300, which has at least one functional well 310 for holding PCR tube caps 920 and test tubes, including reagent tubes and PCR tubes 910. Both the first temperature control mechanism 400 and the second temperature control mechanism 500 are used to regulate the temperature of the items stored in the functional well 310 of the reagent kit 300. The temperature control ranges of the first temperature control mechanism 400 and the second temperature control mechanism 500 are different.

[0039] This setup can meet the diverse needs of routine nucleic acid testing and methylation. It eliminates the need to transfer PCR tubes 910 and other items to a dedicated temperature-controlled device for methylation when required, improving the efficiency of the reagent detection system 10 and avoiding the risk of sample contamination caused by frequent opening, closing, and transferring of PCR tubes 910 and other items.

[0040] In this embodiment, the first temperature control mechanism 400 has a temperature control range of 0-105℃, and can heat or cool the items stored in the functional wells 310 of the reagent kit 300. The second temperature control mechanism 500 has a temperature control range of 25℃ (room temperature)-105℃, and only heats the reagent kit 300 and the items stored in the functional wells 310.

[0041] In another embodiment, the first temperature control mechanism 400 only cools the items stored in the functional wells 310 of the reagent kit 300, and the second temperature control mechanism 500 only heats the reagent kit 300 and the items stored in the functional wells 310.

[0042] The reagent testing system 10 also includes a control device; the control device is electrically connected to the robotic arm device 12, centrifuge device 13, PCR testing device 14, nucleic acid extraction device 15, sample processing device 16, and PCR reaction device 17 for control purposes. The control device includes a power supply unit, a human-machine interface, a communication unit, a processor, and a control unit. The power supply unit can be an external power source or a battery. The human-machine interface can be a computer input / output device such as a display screen, keyboard, touchscreen, buttons, knobs, speakers, and LEDs, used for inputting commands and reading information, thereby achieving human-machine interaction and information exchange. The communication unit can be a transceiver, and the control unit can be a microcontroller unit (MCU).

[0043] Please refer to Figure 3This is a schematic diagram of the PCR reaction apparatus 17 shown in one embodiment of this application. A first temperature control mechanism 400 is disposed on the frame 200 and fixed to the bottom surface of the reagent kit 300, used for temperature control of the entire reagent kit 300. The first temperature control mechanism 400 includes a semiconductor cooling chip, a fan, and an air outlet duct. In another embodiment, the first temperature control mechanism 400 may only include a fan, disposed on the frame 200, for cooling the reagent kit 300 by blowing air onto it. In another embodiment, the first temperature control mechanism 400 may only include a semiconductor cooling chip, disposed on the reagent kit 300, for temperature control of the entire reagent kit 300.

[0044] The second temperature control mechanism 500 is located above the reagent kit 300. The second temperature control mechanism 500 includes a temperature regulating element 510 and at least one temperature conducting element 520. The temperature conducting element 520 is made of a heat-resistant material, such as metal. The temperature regulating element 510 can be a heating block, a heating film, or a semiconductor cooling chip, used to regulate the temperature of the temperature conducting element 520. The PCR reaction apparatus 17 also includes a motion mechanism 600. The motion mechanism 600 is located on the frame 200 and is connected to the temperature conducting element 520 for driving the temperature conducting element 520 closer to or further away from the functional well position 310.

[0045] With this configuration, the motion mechanism 600 can drive the temperature-conducting element 520 to reach the designated functional port 310, thereby heating individual designated consumables in the reagent kit 300.

[0046] In this embodiment, the temperature regulating component 510 and the temperature guiding component 520 are directly fixed together by means of bolts or other methods, and the motion mechanism 600 drives the temperature guiding component 520 and the temperature regulating component 510 to move together at the same time.

[0047] In another embodiment, the temperature regulating element 510 and the temperature conducting element 520 can be detachably connected together by means of snap-fit ​​or similar means, or the temperature regulating element 510 can be directly fixed to the frame 200, and the temperature of the temperature conducting element 520 can be raised through contact. The motion mechanism 600 only drives the temperature conducting element 520 to move, without moving the temperature regulating element 510. For example, the motion mechanism 600 drives the temperature conducting element 520 to the temperature regulating element 510, so that the temperature conducting element 520 can move to the temperature regulating element 510 for heating, and then the motion mechanism 600 drives the temperature conducting element 520 to move to other functional positions 310 to heat the test tube and the reagents inside the test tube.

[0048] The functional well positions 310 are provided in multiple ways, including at least one cap storage hole 311 for placing a PCR tube cap 920 and at least one tube storage hole 312 for placing test tubes (e.g., reagent tubes and PCR tubes 910); wherein, the temperature conducting element 520 is configured to connect with the test tube cap (e.g., PCR tube cap 920) to drive the PCR tube cap 920 to move.

[0049] The temperature-conducting component 520 has a cylindrical structure and can be detachably connected to the PCR tube cap 920 via snap-fit ​​or plug-in methods. The temperature-conducting component 520 can be cylindrical, cuboid, or pentagonal prism.

[0050] PCR reaction apparatus 17 also includes: cap removal mechanism 800, which is connected to motion mechanism 600 and is used to separate temperature conducting element 520 from PCR tube cap 920.

[0051] With this configuration, the temperature-conducting component 520 of the second temperature control mechanism 500 in this embodiment can be connected to the PCR tube cap 920 to drive the PCR tube cap 920 to move, thereby realizing the automatic opening and closing of the PCR tube 910 and transferring the PCR tube cap 920. This achieves automation, reduces manual intervention, and eliminates the need for the reagent detection system 10 to have a separate opening and closing mechanism for the PCR tube 910. This simplifies the overall structure of the reagent detection system 10, making it more compact and reducing costs.

[0052] In another embodiment, the temperature-conducting element 520 can directly engage with the PCR tube 910 without passing through the PCR tube cap 920, thereby heating the PCR tube 910 and the reagents inside the tube.

[0053] In this embodiment, the test tube placed in the functional well position 310 includes a row of tubes composed of multiple interconnected PCR tubes 910. Multiple temperature-conducting elements 520 are provided, arranged in a linear array on the heating block of the temperature-regulating element 510. These elements can cooperate with multiple PCR tubes 910 or multiple PCR tube caps 920 simultaneously to heat the row of tubes. The number of temperature-conducting elements 520 can be 8, 24, or 96.

[0054] In another embodiment, the temperature conductor 520 is provided to cooperate with a single PCR tube 910 or a single PCR tube cap 920 to achieve independent heating of the single PCR tube 910.

[0055] Please refer to Figure 4 This is an exploded schematic diagram of the PCR reaction apparatus 17 shown in an embodiment of this application. The distribution of the multiple storage tube holes 312 can be divided into a PCR tube area 301, a reagent tube area 302, and a sample retention tube area 303; wherein, the PCR tube area 301 and the reagent tube area 302 are located on the left and right sides of the multiple storage cap holes 311, respectively, and the sample retention tube area 303 is located between the reagent tube area 302 and the storage cap holes 311.

[0056] Since the storage well 312 in the reagent tube area 302 can be used to store reagents that require refrigeration, and the storage well 312 in the sample retention tube area 303 can be used to store samples, while the PCR tube 910 in the PCR tube area 301 may be heated by the second temperature control mechanism 500, the PCR tube area 301 and the reagent tube area 302 are located on both sides of the multiple storage wells 311, and the sample retention tube area 303 is located between the reagent tube area 302 and the storage wells 311. The multiple storage wells 311 can play a certain role in heat insulation, so as to prevent the heating of the PCR tube 910 by the second temperature control mechanism 500 from affecting the samples or reagents in the reagent tube area 302 and the sample retention tube area 303.

[0057] The PCR reaction apparatus 17 also includes a clamping element 700 and a clamping element drive mechanism 730. The clamping element 700 is configured to approach or move away from the functional aperture 310. The clamping element drive mechanism 730 is located on the frame 200 and is connected to the clamping element 700 for driving the clamping element 700. The clamping element drive mechanism 730 can be a cylinder, a lead screw motor, or other components.

[0058] With this configuration, the clamping component 700 can clamp the test tube, preventing the test tube or PCR tube cap 920 from leaving the reagent kit 300 under the drive of the temperature-conducting component 520.

[0059] Please refer to Figure 5 This is an exploded view of a PCR reaction apparatus 17 according to an embodiment of this application. The clamping member 700 includes a main body plate 710, on which at least one pressure rod 720 is connected. The main body plate 710 is configured to be pushed by the temperature-conducting member 520. With this configuration, the movement of the main body plate 710 can be indirectly controlled by controlling the movement of the temperature-conducting member 520 through the motion mechanism 600, thereby eliminating the need for a separate clamping member drive mechanism 730, simplifying the structure of the entire PCR reaction apparatus 17, and reducing costs.

[0060] The temperature-conducting component 520 can push the main body plate 710 in various ways. For example, the main body plate 710 can be pushed by the temperature-conducting component 520 directly contacting the side surface of the main body plate 710; or the main body plate 710 and the temperature-conducting component 520 can be detachably connected by providing mutually cooperating buckles and grooves 911 on the main body plate 710 and the temperature-conducting component 520 respectively, so as to push the main body plate 710.

[0061] In this embodiment, a drive groove 711 is provided on the main body plate 710 for the temperature-conducting component 520 to extend into. After the temperature-conducting component 520 extends into the drive groove 711, it can push the main body plate 710. The drive groove 711 can be an elongated hole into which multiple temperature-conducting components 520 can extend.

[0062] In the bundled tubes, each PCR tube 910 has a groove 911 on its outer wall, and a connecting rod 912 is provided between two adjacent PCR tubes 910. A clamping rod 720 is configured to extend into the groove 911 and press against the connecting rod 912. This configuration allows the bundled tubes to be clamped together by the clamping rod 720, preventing them from leaving the kit 300 due to the movement of the temperature-conducting element 520. The number of clamping rods 720 corresponds to the number of PCR tubes 910 in the bundled tubes; for example, the difference between the number of clamping rods 720 and the number of PCR tubes 910 in the bundled tubes is 1.

[0063] The PCR reaction apparatus 17 further includes a first slide rail 740 and a first slider 750. The first slide rail 740 is disposed on the frame 200 and located on one side of the reagent kit 300. The first slider 750 is slidably disposed on the first slide rail 740 and connected to the main plate 710. The arrangement of the first slide rail 740 and the first slider 750 allows the main plate 710 to be movably disposed on the frame 200, and also restricts the direction of movement and vertical position of the main plate 710. Two of each of the first slide rail 740 and the first slider 750 are provided.

[0064] The motion mechanism 600 includes a translation mechanism 630, a lifting mechanism 620, and a mounting base 610. The mounting base 610 is movably mounted above the frame 200 via a second slide rail 640 and a second slider 650. There are two second slide rails 640 and two second sliders 650, with two first slide rails 740 positioned between the two second slide rails 640. The lifting mechanism 620 includes a lifting motor 621 and a belt pulley transmission mechanism 622, both mounted on the mounting base 610. The lifting motor 621 is connected to the temperature-conducting component 520 via the belt pulley transmission mechanism 622, driving the temperature-conducting component 520 to rise and fall.

[0065] The translation mechanism 630 includes components such as a lead screw motor or a cylinder. The translation mechanism 630 is located on the frame 200 and is connected to the mounting base 610 for driving the mounting base 610 to translate along the direction of the second slide rail 640.

[0066] In addition, a temperature sensor 530 and an overheat switch 540 are installed on the temperature control component 510 through holes or slots, and the temperature sensor 530 and the overheat switch 540 are electrically connected to the temperature control component 510.

[0067] During operation, the temperature regulating element 510 is energized for heating. The temperature sensor 530 senses the real-time temperature and sends it to the control device. The control device controls the human-machine interface to display the temperature information in real time and determines whether the temperature regulating element 510 has reached the set temperature (e.g., within ±1 degree). If so, the control device controls the temperature regulating element 510 to heat intermittently. During the heating process of the temperature regulating element 510, the overheat switch 540 implements overheat protection. The triggering principle of the overheat switch 540 is that after overheating, the deformation of the two contacts in the overheat switch 540 exceeds the range, and the two contacts disconnect, without requiring electrical logic judgment.

[0068] Please refer to Figure 6 This is a front view of a PCR reaction apparatus 17 shown in one embodiment of this application. The temperature-conducting element 520 is solid, and the PCR tube cap 920 may have a connecting groove 921 that matches the temperature-conducting element 520. The temperature-conducting element 520 and the PCR tube 910 are detachably connected by an insertion method. In another embodiment, the temperature-conducting element 520 has a slot, and the PCR tube cap 920 may have a protrusion that matches the slot.

[0069] Among them, each of the two outermost temperature-conducting components 520 is provided with a slot 521, and each of the two outermost connecting slots 921 on the PCR tube cap 920 is provided with a spring piece 922 that matches the slot 521.

[0070] The cap removal mechanism 800 includes at least one cap removal plate 801, which is fixed to the mounting base 610 and arranged along the axis of the temperature-conducting element 520. When the temperature-conducting element 520 and the cap removal plate 801 move relative to each other, the cap removal mechanism 800 extends through the cap removal plate 801 between the slot 521 and the spring clip 922, thereby separating the temperature-conducting element 520 from the PCR tube cap 920. In this embodiment, two cap removal plates 801 are provided, each corresponding to one of the two slots 521.

[0071] There are several ways to achieve relative movement between the temperature-conducting component 520 and the cover-removing plate 801. For example, the lifting mechanism 620 can drive the temperature-conducting component 520 to move up and down, causing the temperature-conducting component 520 to move up and down relative to the cover-removing plate 801. Alternatively, a moving mechanism can be provided in the cover-removing mechanism 800 to drive the cover-removing plate 801 to move up and down. The moving mechanism includes a cylinder or a lead screw motor. The cover-removing plate 801 can be moved up and down relative to the temperature-conducting component 520 through the moving mechanism.

[0072] Please refer to Figure 4 and Figure 5During operation, the translation mechanism 630 drives the mounting base 610 and moves the temperature-conducting component 520 to the clamping component 700. The translation mechanism 630 drives the mounting base 610 and moves the temperature-conducting component 520 into the drive groove 711 of the clamping component 700. The translation mechanism 630 drives the temperature-conducting component 520 to move so as to move the clamping component 700, so that the pressure rod 720 of the clamping component 700 extends into the groove 911 and presses against the connecting rod 912, thereby completing the operation of clamping the connecting pipe and restricting the vertical movement of the connecting pipe.

[0073] After the operation of pressing the tube bundle is completed, the translation mechanism 630 drives the mounting base 610 and moves the temperature-conducting component 520 to the cap storage hole 311. The lifting mechanism 620 drives the temperature-conducting component 520 to move down, and the temperature-conducting component 520 is inserted into the connecting groove 921 of the PCR tube cap 920. At the same time, the spring clip 922 on the PCR tube cap 920 is embedded in the slot 521 of the temperature-conducting component 520, so that the temperature-conducting component 520 is fixedly connected to the PCR tube cap 920. The PCR tube cap 920 can move together with the temperature-conducting component 520 to complete the cap removal operation.

[0074] After the cap removal operation is completed, the lifting mechanism 620 drives the temperature-conducting component 520 and moves the PCR tube cap 920 upward. The translation mechanism 630 drives the mounting base 610 and moves the temperature-conducting component 520 and the PCR tube cap 920 horizontally to the PCR tube area 301. The lifting mechanism 620 then drives the temperature-conducting component 520 and moves the PCR tube cap 920 downward, and the PCR tube cap 920 is repositioned on the PCR tube 910, completing the capping operation.

[0075] After the capping operation is completed, the temperature-conducting element 520 can be heated by the second temperature control mechanism 500. The temperature of the temperature-conducting element 520 is transferred to the reagent in the PCR tube 910 to carry out the heating and conversion operation.

[0076] After the heating and conversion operation is completed, the lifting mechanism 620 drives the temperature-conducting component 520 and moves the PCR tube cap 920 upward. At this time, since the clamping component 700 clamps the tube array, the PCR tube 910 and the PCR tube cap 920 of the tube array can be separated under the action of the lifting mechanism 620, thus completing the opening operation of the PCR tube 910.

[0077] After the PCR tube 910 is opened, the translation mechanism 630 drives the mounting base 610 and moves the temperature-conducting component 520 to the cap storage hole 311. The lifting mechanism 620 drives the temperature-conducting component 520 and moves the PCR tube cap 920 upward, so that the temperature-conducting component 520 and the cap removal plate 801 move relative to each other, so that the cap removal plate 801 extends between the slot 521 and the spring piece 922, so that the temperature-conducting component 520 and the PCR tube cap 920 are separated. The PCR tube cap 920 falls to the cap storage hole 311, completing the cap removal operation.

[0078] Please refer to Figures 1-6 The following is a method of using the reagent detection system 10 for nucleic acid detection, including the following steps:

[0079] Step S101: The reagent detection system 10 is turned on, and the first temperature control mechanism 400 automatically controls the temperature of the entire reagent kit 300 to 4℃.

[0080] Step S102: The sample tubes in the sample holder are processed by the sample processing device 16 and the first robotic arm 121, including cupping and opening. At the same time, extraction reagents (lysis buffer, washing buffer, elution buffer, magnetic beads, etc.) are added to the extraction consumables located at the nucleic acid extraction device 15 by the pipette of the second robotic arm 122.

[0081] Step S103: Add the sample from the sample tube to the extraction consumables located at the nucleic acid extraction device 15 using the pipette of the second robotic arm 122.

[0082] Step S104: Nucleic acid extraction (lysis, washing, elution) is performed using nucleic acid extraction device 15.

[0083] Step S105: After extraction, the extracted nucleic acid product is transferred to the PCR tube 910 in the PCR tube area 301 of the PCR tube 910 using the pipette of the second robotic arm 122.

[0084] Step S106: Use the pipette of the second robotic arm 122 to put primers, protease and other PCR reagents into the PCR tube 910 to construct the system.

[0085] Step S107: The second temperature control mechanism 500 completes the cap removal, cap pressing, and cap unsealing operations.

[0086] Step S108: The capped PCR tube 910 is transferred to the centrifuge device 13 by the third robotic arm 123 for centrifugation.

[0087] Step S109: After centrifugation, the PCR tube 910 is transferred to the PCR detection device 14 for detection by the third robotic arm 123.

[0088] Please refer to Figures 1-6 The following is a method of using the reagent detection system 10 for performing methylation experiments, including the following steps:

[0089] Step S201: The reagent detection system 10 is turned on, and the first temperature control mechanism 400 automatically controls the temperature of the entire reagent kit 300 to 4°C.

[0090] Step S202: The sample tubes in the sample holder are processed by the sample processing device 16 and the first robotic arm 121, including dispensing and opening the caps. At the same time, extraction reagents (lysis buffer, washing buffer, elution buffer, magnetic beads, etc.) are added to the extraction consumables located at the nucleic acid extraction device 15 by the pipette of the second robotic arm 122.

[0091] Step S203: Add the sample from the sample tube to the extraction consumables located at the nucleic acid extraction device 15 using the pipette of the second robotic arm 122.

[0092] Step S204: Perform the first nucleic acid extraction (lysis, washing, elution) using the nucleic acid extraction device 15.

[0093] Step S205: After extraction, the extracted nucleic acid product is transferred to the PCR tube 910 in the PCR tube area 301 of the PCR tube 910 using the pipette of the second robotic arm 122.

[0094] Step S206: Using the pipette of the second robotic arm 122, water and conversion reagent (sodium bisulfite) and other reagents are added to the PCR tube 910 to construct the system.

[0095] Step S207: The cap removal and cap pressing operations are completed by the second temperature control mechanism 500.

[0096] Step S208: The PCR tube 910 is heated and transformed by the second temperature control mechanism 500 (Heating process example: 95℃ 5min → 60℃ 20min → 95℃ 5min → 60℃ 20min).

[0097] Step S209: After the heating and conversion operation is completed, the second temperature control mechanism 500 is used to open and uncap the PCR tube 910.

[0098] Step S210: The methylated product in the PCR tube 910 is transferred to the nucleic acid extraction device 15 by the pipette of the second robotic arm 122.

[0099] Step S211: Perform a second nucleic acid extraction using the nucleic acid extraction device 15 (perform 4 washes and 1 elution; the difference from the first nucleic acid extraction is that lysis is not required). At the same time, the first temperature control mechanism 400 operates (or starts cooling immediately upon startup) to apply low temperature to the PCR tube area 301.

[0100] Step S212: After the temperature of the first temperature control mechanism 400 stabilizes at 4℃, put primers, protease and other reagents into PCR tube 910.

[0101] Step S213: After the second nucleic acid extraction is completed, the second nucleic acid extraction product is transferred back to the PCR automated temperature control device for system construction using the pipette of the second robotic arm 122 (PCR reagents are added into PCR tube 910).

[0102] Step S214: The cap removal, cap pressing, and cap unsealing operations are completed by the second temperature control mechanism 500.

[0103] Step S215: The capped PCR tube 910 is transferred to the centrifuge device 13 by the third robotic arm 123 for centrifugation.

[0104] Step S216: After centrifugation, the PCR tube 910 is transferred to the PCR detection device 14 for detection by the third robotic arm 123.

[0105] It should be noted that, where there is no conflict, the features in the embodiments of this application can be combined with each other. The above are merely preferred embodiments of this application and are not intended to limit this application. For those skilled in the art, this application can have various modifications and variations. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of this application should be included within the protection scope of this application.

Claims

1. A PCR reaction apparatus, characterized in that, include: frame; A reagent kit is disposed on the rack, and the reagent kit has at least one functional well. A first temperature control mechanism is located on the frame; A second temperature control mechanism is located above the reagent kit. The second temperature control mechanism includes a temperature regulating element and at least one temperature conducting element. The temperature regulating element is used to adjust the temperature of the temperature conducting element. A motion mechanism, located on the frame and connected to the temperature-conducting component, is used to drive the temperature-conducting component closer to or further away from the functional hole. A clamping member, configured to approach or move away from the functional aperture, the clamping member comprising: A main body plate, the main body plate being configured to be pushed by the heat-conducting element; At least one pressure bar is connected to the main body plate; The functional ports are provided for placing test tubes. The test tubes include a row of tubes composed of multiple interconnected PCR tubes. Each PCR tube has a groove on its outer wall, and a connecting rod is provided between two adjacent PCR tubes. The pressure rod is configured to extend into the groove and press against the connecting rod. The main body plate is provided with a drive groove for the temperature-conducting component to extend into. The plurality of said functional holes also include: at least one cap storage hole for placing a test tube cap and at least one tube storage hole for placing a test tube; The temperature-conducting element is configured to connect with the test tube cap to move the test tube cap.

2. The PCR reaction apparatus according to claim 1, characterized in that, The PCR reaction apparatus further includes: A clamping member drive mechanism is located on the frame and is connected to the clamping member for driving the clamping member to move.

3. The PCR reaction apparatus according to claim 1, characterized in that, The PCR reaction apparatus further includes: A first slide rail is disposed on the rack and located on one side of the reagent kit; and The first slider is slidably disposed on the first slide rail and connected to the main body plate.

4. The PCR reaction apparatus according to claim 1, characterized in that, The PCR reaction apparatus further includes: A cap removal mechanism, located within the motion mechanism, is used to separate the temperature-conducting element from the test tube cap.

5. The PCR reaction apparatus according to claim 4, characterized in that, The motion mechanism includes: The mounting base is located above the frame; A lifting mechanism is provided on the mounting base and is connected to the temperature-conducting component for driving the temperature-conducting component to rise and fall; A translation mechanism is provided on the frame and is connected to the mounting base for driving the mounting base to translate. The cover removal mechanism includes at least one cover removal plate, which is fixed to the mounting base and arranged along the axis of the temperature-conducting component.

6. A reagent detection system, characterized in that, include: Chassis; The robotic arm device is located inside the chassis; as well as The PCR reaction apparatus as described in any one of claims 1 to 5 is disposed within the chassis.

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