PCR (Polymerase Chain Reaction) clamping box device

Abstract

The utility model discloses a PCR reaction cartridge device, which comprises a PCR reaction chamber, a hybridization reaction chamber, a first flow channel, a second flow channel, a third flow channel, a fourth flow channel, a PCR sample adding port, a hybridization reagent port and a thrust inlet, the hybridization reaction chamber is communicated with the PCR reaction chamber, and the center position of the bottom of the hybridization reaction chamber is provided with a discharge port communicated with the outside; one end of the first runner is communicated with the PCR reaction bin, the other end of the first runner is communicated with the hybridization reaction bin, and a first opening and closing assembly is arranged on the first runner; one end of the second flow channel is communicated with the hybridization reaction bin, and a second opening and closing assembly is arranged on the second flow channel; one end of the third flow channel is communicated with the PCR reaction bin, and a third opening and closing assembly is arranged on the third flow channel; one end of the fourth flow channel is communicated with the third flow channel between the third opening and closing assembly and the PCR reaction bin, and a fourth opening and closing assembly is arranged on the fourth flow channel. Through the arrangement, the PCR reaction bin and the hybridization reaction bin are integrated, so that the operation process is simplified, the pollution risk is reduced, and the experiment effect is improved.

Description

Technical Field

[0001] This application relates to the technical field of nucleic acid amplification and hybridization equipment, and particularly relates to a PCR reaction cartridge device. Background Art

[0002] Polymerase Chain Reaction (PCR) is a molecular biology technique used to amplify specific DNA fragments. PCR consists of three basic reaction steps: denaturation - annealing - extension. Among them, denaturation means that the template DNA is heated to 94 - 98 °C to dissociate the DNA double strand or the double-stranded DNA formed by PCR amplification, turning the template DNA into a single-stranded template. Annealing means cooling the single strand formed during the denaturation process to 50 - 65 °C to allow the primer to pair and bind with the complementary sequence of the template DNA single strand. Extension means that at a relatively high temperature (usually around 72 °C), the DNA template and primer conjugate synthesize a new semi-conservative replication strand complementary to the template DNA strand according to the base pairing and semi-conservative replication principle. Repeating the above three steps of denaturation - annealing - extension can obtain more semi-conservative replication strands, and this new strand can become the template for the next cycle, thereby ultimately achieving the amplification of DNA fragments.

[0003] Reverse Dot Blot (RDB) technology is to first spot the probes to be used onto a nitrocellulose membrane or nylon membrane in sequence, and then hybridize the DNA sample to be tested (usually the product specifically amplified by PCR, with biotin labeled at the PCR primer end in advance, so that the amplification product is correspondingly labeled with biotin) with the probes. In this way, the amplified product of the sample to be tested will specifically bind to the probe with a homologous sequence, and the unbound product DNA sample is removed by washing. Since the DNA sample to be tested has a biotin-like marker, the probe spot bound to the DNA to be tested will carry a biotin-like marker, and then the hybridization signal can be shown through the corresponding color reaction. In this way, multiple targets in the sample can be screened simultaneously in a single reaction, saving time, effort, and reagent consumables. Due to its low hardware cost and convenient result reading, RDB technology has been widely used in the field of medical diagnosis, such as genotyping detection, genetic disease detection, pathogen detection, tumor gene detection, etc.

[0004] In a molecular laboratory, PCR and hybridization reactions are commonly used key technologies for the amplification and detection of specific DNA or RNA sequences. However, in the current experimental process, PCR and hybridization reactions usually require the use of different reaction cartridges or devices. Operators need to frequently switch between the two reactions, which is cumbersome and difficult. Using different reaction devices will increase the risk of experimental contamination. At the same time, it is prone to being affected by contamination, resulting in unstable experimental results. Therefore, there is an urgent need to design a reaction cartridge device that integrates PCR and hybridization methods to solve the above problems. Summary of the Utility Model

[0005] The main purpose of this application is to provide a PCR reaction cartridge device, aiming to solve the problems of cumbersome operation process, high contamination risk, and unstable experimental results in existing PCR and hybridization reaction cartridges.

[0006] To achieve the above object, this application provides a PCR reaction cartridge device, including a PCR reaction chamber, a hybridization reaction chamber, a first flow channel, a second flow channel, a third flow channel, a fourth flow channel, a PCR sample addition port, a hybridization reagent port, and a thrust inlet. Among them, the hybridization reaction chamber is communicated with the PCR reaction chamber, and a discharge port communicating with the outside is provided at the center of the bottom of the hybridization reaction chamber; one end of the first flow channel is communicated with the PCR reaction chamber and the other end is communicated with the hybridization reaction chamber, and a first opening and closing component is provided on the first flow channel; one end of the second flow channel is communicated with the hybridization reaction chamber, and a second opening and closing component is provided on the second flow channel; one end of the third flow channel is communicated with the PCR reaction chamber, and a third opening and closing component is provided on the third flow channel; one end of the fourth flow channel is communicated with the third flow channel between the third opening and closing component and the PCR reaction chamber, and a fourth opening and closing component is provided on the fourth flow channel; the PCR sample addition port is communicated with the other end of the third flow channel; the hybridization reagent port is communicated with the other end of the second flow channel; the thrust inlet is communicated with the other end of the third flow channel.

[0007] Optionally, the PCR reaction cartridge device further includes: a top cover and a base. Among them, a groove is provided on the surface of the base close to the top cover, and the base is bonded to the top cover so that the groove forms the PCR reaction chamber, the hybridization reaction chamber, and the first flow channel, the second flow channel, the third flow channel, and the fourth flow channel.

[0008] Optionally, the first opening and closing component, the second opening and closing component, the third opening and closing component, and the fourth opening and closing component all adopt a membrane pump structure.

[0009] Optionally, the top cover includes: a PCR sample addition tube, a hybridization reagent tube, four pressure holes, and a thrust hole. Among them, the PCR sample addition tube is arranged at the position of the top cover corresponding to the PCR sample addition port, and one end of the PCR sample addition tube penetrates through the top cover and communicates with the PCR sample addition port, and the other end communicates with the outside; the hybridization reagent tube is arranged at the position of the top cover corresponding to the hybridization reagent port, and one end of the hybridization reagent tube penetrates through the top cover and communicates with the hybridization reagent port, and the other end communicates with the outside; the four pressure holes are respectively opened at the positions of the top cover corresponding to the first opening and closing assembly, the second opening and closing assembly, the third opening and closing assembly, and the fourth opening and closing assembly; the thrust hole is opened at the position of the top cover corresponding to the thrust inlet.

[0010] Optionally, a discharge tube is provided at the position of the base corresponding to the discharge port, one end of the discharge tube communicates with the discharge port, and the other end communicates with the outside.

[0011] Optionally, the PCR reaction cartridge device further includes a PC film, the PC film is located between the top cover and the base, and through holes are respectively provided at the positions of the PC film corresponding to the PCR sample addition port, the hybridization reagent port, and the thrust inlet.

[0012] Optionally, a pressure - sensitive adhesive is coated on the surface of the PC film where it contacts the top cover and the base.

[0013] Optionally, an observation port is provided at the position of the top cover corresponding to the hybridization reaction chamber, and a transparent plate is fixedly installed at the observation port.

[0014] Optionally, a membrane chip is provided in the hybridization reaction chamber.

[0015] Optionally, the inner bottom surface of the hybridization reaction chamber is an inclined surface.

[0016] A PCR reaction cartridge device proposed in an embodiment of the present application integrates a PCR reaction chamber and a hybridization reaction chamber into the same device to simplify the operation process of molecular experiments. The settings of the first flow channel, the second flow channel, the third flow channel, the fourth flow channel, and the first opening and closing assembly, the second opening and closing assembly, the third opening and closing assembly, and the fourth opening and closing assembly can control the sequence of reactions, which is convenient for operation. At the same time, one end of the fourth flow channel is provided with a PCR sample addition port, one end of the second flow channel is provided with a hybridization reagent port, and one end of the third flow channel is provided with a thrust inlet, realizing the operations of PCR and hybridization reactions in a closed device, reducing the risk of aerosol contamination, and greatly improving the experimental effects of PCR and hybridization reactions through the above - mentioned settings. Description of the Drawings

[0017] Figure 1 It is a schematic structural diagram of the PCR reaction cartridge device provided by the embodiment of the present application;

[0018] Figure 2 This is a schematic exploded view of the PCR reaction cassette device provided by the embodiments of the present application;

[0019] Figure 3 is Figure 1 a schematic cross-sectional view of the PCR reaction cassette device shown along the A-A' direction;

[0020] Figure 4 is Figure 2 a top view of the base of the PCR reaction cassette device shown.

[0021] Reference numerals: 1, base; 101, first flow channel; 102, second flow channel; 103, third flow channel; 104, fourth flow channel; 105, PCR sample addition port; 106, hybridization reagent port; 107, thrust inlet; 11, PCR reaction chamber; 12, hybridization reaction chamber; 13, discharge port; 14, first opening and closing assembly; 15, second opening and closing assembly; 16, third opening and closing assembly; 17, fourth opening and closing assembly; 18, discharge pipe; 2, top cover; 21, PCR sample addition tube; 22, hybridization reagent tube; 23, four pressure holes; 24, thrust hole; 25, observation port; 3, PC film.

[0022] The realization, functional features and advantages of the objectives of the present application will be further described in conjunction with the embodiments with reference to the accompanying drawings. Detailed implementation manners

[0023] Next, the technical solutions in the embodiments of the present application will be clearly and completely described in conjunction with the accompanying drawings in the embodiments of the present application. Obviously, the described embodiments are only a part of the embodiments of the present application, rather than all the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments in the present application without making creative efforts shall fall within the protection scope of the present application.

[0024] It should be noted that all directional indications (such as up, down, left, right, front, back...) in the embodiments of the present application are only used to explain the relative positional relationship and movement conditions between components in a specific posture (as shown in the accompanying drawings). If the specific posture changes, the directional indications will also change accordingly.

[0025] In this application, unless otherwise clearly defined and limited, terms such as "connection" and "fixation" shall be understood in a broad sense. For example, "fixation" can be a fixed connection, a detachable connection, or integrated; it can be a mechanical connection or an electrical connection; it can be directly connected or indirectly connected through an intermediate medium, and can be the communication inside two components or the interaction relationship between two components, unless otherwise clearly limited. For those of ordinary skill in the art, the specific meanings of the above terms in this application can be understood according to specific circumstances.

[0026] In addition, if there are descriptions involving "first", "second", etc. in the embodiments of this application, the descriptions of "first", "second", etc. are only for descriptive purposes and cannot be understood as indicating or implying their relative importance or implicitly indicating the quantity of the indicated technical features. Thus, the features defined with "first" and "second" can explicitly or implicitly include at least one of such features. In addition, the meaning of "and / or" appearing throughout the text includes three parallel scenarios. Taking "A and / or B" as an example, it includes scenario A, or scenario B, or the scenario where A and B are satisfied simultaneously. In addition, the technical solutions between various embodiments can be combined with each other, but it must be based on the fact that those of ordinary skill in the art can implement them. When the combination of technical solutions results in contradictions or cannot be implemented, it should be considered that such a combination of technical solutions does not exist and is not within the scope of protection required by this application.

[0027] The following will describe this application in detail with reference to the accompanying drawings and embodiments.

[0028] Figure 1 It is a schematic structural diagram of the PCR reaction cartridge device provided by the embodiment of this application. Figure 2 It is an exploded structural diagram of the PCR reaction cartridge device provided by the embodiment of this application. Figure 3 is Figure 1 The cross-sectional structural diagram of the PCR reaction cartridge device shown along the A-A' direction. Figure 4 is Figure 2 The top view of the base of the PCR reaction cartridge device shown.

[0029] Please refer to Figure 1 , Figure 2 and Figure 4Referring to this, an embodiment of the present application provides a PCR reaction cartridge device, which may include a PCR reaction chamber 11, a hybridization reaction chamber 12, a first flow channel 101, a second flow channel 102, a third flow channel 103, a fourth flow channel 104, a PCR sample addition port 105, a hybridization reagent port 106, and a thrust inlet 107; wherein, the hybridization reaction chamber 12 is communicated with the PCR reaction chamber 11, and a discharge port 13 communicating with the outside is provided at the center of the bottom surface of the hybridization reaction chamber 12; one end of the first flow channel 101 is communicated with the PCR reaction chamber 11 and the other end is communicated with the hybridization reaction chamber 12, and a first opening and closing component 14 is provided on the first flow channel 101; one end of the second flow channel 102 is communicated with the hybridization reaction chamber 12, and a second opening and closing component 15 is provided on the second flow channel 102; one end of the third flow channel 103 is communicated with the PCR reaction chamber 11, and a third opening and closing component 16 is provided on the third flow channel 103; one end of the fourth flow channel 104 is communicated with the third flow channel 103 between the third opening and closing component 16 and the PCR reaction chamber 11, and a fourth opening and closing component 17 is provided on the fourth flow channel 104; the PCR sample addition port 105 is communicated with the other end of the fourth flow channel 104; the hybridization reagent port 106 is communicated with the other end of the second flow channel 102; the thrust inlet 107 is communicated with the other end of the third flow channel 103.

[0030] In this embodiment, the three basic reaction steps of "denaturation - annealing - extension" of PCR are carried out in the PCR reaction chamber 11 to achieve the amplification of DNA fragments.

[0031] When using the above - mentioned PCR reaction cartridge device, the following steps can be carried out:

[0032] S1. Open the fourth opening and closing component 17 and the first opening and closing component 14, add a sample and a PCR reaction solution to the PCR sample addition port 105. The sample and the PCR reaction solution sequentially enter the PCR reaction chamber 11 through the fourth flow channel 104 and the third flow channel 103. After the sample addition is completed, close the fourth opening and closing component 17 and the first opening and closing component 14.

[0033] S2. Through an external temperature control device, control the temperature of the PCR reaction chamber 11 to complete the three basic reaction steps of "denaturation - annealing - extension" of PCR.

[0034] S3. After completing the above - mentioned step S2 (i.e., the PCR amplification reaction), open the third opening and closing component 16 and the first opening and closing component 14. Through the push of an external thrust force via the thrust inlet 107, the reaction solution in the PCR reaction chamber 11 enters the hybridization reaction chamber 12 through the first flow channel 101. After the reaction solution completely enters the hybridization reaction chamber 12, close the third opening and closing component 16 and the first opening and closing component 14.

[0035] S4. Open the second opening and closing component 15, and sequentially add hybridization solution, washing solution, enzyme-labeled solution, washing solution, chromogenic solution, and washing solution through the hybridization reagent port 106; the hybridization solution, washing solution, enzyme-labeled solution, washing solution, chromogenic solution, and washing solution enter the hybridization reaction chamber 12 through the second flow channel 102 (it should be noted that the waste liquid generated in this step S4 is discharged from the discharge port 13). The hybridization, enzyme-labeling, chromogenic, and washing processes are completed in the hybridization reaction chamber 12.

[0036] A PCR reaction cassette device proposed in an embodiment of the present application integrates the PCR reaction chamber 11 and the hybridization reaction chamber 12 into the same device to simplify the operation process of molecular experiments. The settings of the first flow channel 101, the second flow channel 102, the third flow channel 103, the fourth flow channel 104, and the first opening and closing component 14, the second opening and closing component 15, the third opening and closing component 16, and the fourth opening and closing component 17 can control the sequence of reactions, which is convenient for operation. At the same time, one end of the fourth flow channel 104 is provided with a PCR sample addition port, one end of the second flow channel 102 is provided with a hybridization reagent port 106, and one end of the third flow channel 103 is provided with a thrust inlet 107, realizing the operation of PCR and hybridization reactions in a closed device, reducing the risk of aerosol contamination, and greatly improving the experimental effects of PCR and hybridization reactions through the above settings.

[0037] Please refer to Figure 1 and Figure 2 In an exemplary embodiment, the PCR reaction cassette device may further include a top cover 2 and a base 1; wherein, a groove is provided on the surface of the base 1 close to the top cover 2, and the base 1 and the top cover 2 are bonded to form the PCR reaction chamber 11, the hybridization reaction chamber 12, and the first flow channel 101, the second flow channel 102, the third flow channel 103, and the fourth flow channel 104.

[0038] It can be understood that a groove is provided on the surface of the base 1 close to the top cover 2, and the shape and size of the groove correspond to those of the PCR reaction chamber 11, the hybridization reaction chamber 12, and the first flow channel 101, the second flow channel 102, the third flow channel 103, and the fourth flow channel 104.

[0039] In one embodiment, the first opening and closing component 14, the second opening and closing component 15, the third opening and closing component 16, and the fourth opening and closing component 17 can all adopt a membrane pump structure.

[0040] Specifically, through the pump membrane structure, the positive and negative pressures of the membrane are used to control the flow and stop of the reaction solution. At the position where the membrane pump structure is provided on the base 1, the groove is set in a disconnected state. The membrane pump structure can accurately control the flow rate and direction of the reaction solution through pressure adjustment, ensuring the full mixing of the reaction solution and its effective binding with the probe or primer.

[0041] The pump membrane structure can effectively prevent cross - contamination of the reaction solution through a highly sealed design, thereby ensuring the accuracy and reliability of the experimental results.

[0042] It should be noted that in this embodiment, the first opening - closing component 14, the second opening - closing component 15, the third opening - closing component 16, and the fourth opening - closing component 17 can all adopt the membrane pump structure, or any device structure that can achieve the switching of the channel connection or disconnection state.

[0043] In an embodiment, the top cover 2 includes a PCR sample addition tube 21, a hybridization reagent tube 22, four pressure holes 23, and a thrust hole 24; among them, the PCR sample addition tube 21 is arranged at the position of the top cover 2 corresponding to the PCR sample addition port 105, and one end of the PCR sample addition tube 21 penetrates through the top cover 2 and is connected to the PCR sample addition port 105, and the other end is connected to the outside; the hybridization reagent tube 22 is arranged at the position of the top cover 2 corresponding to the hybridization reagent port 106, and one end of the hybridization reagent tube 22 penetrates through the top cover 2 and is connected to the hybridization reagent port 106, and the other end is connected to the outside; the four pressure holes 23 are respectively opened at the positions of the top cover 2 corresponding to the first opening - closing component 14, the second opening - closing component 15, the third opening - closing component 16, and the fourth opening - closing component 17; the thrust hole 24 is opened at the position of the top cover 2 corresponding to the thrust inlet 107.

[0044] Specifically, the settings of the PCR sample addition tube 21 and the hybridization reagent tube 22 play a role in guiding the flow, so that the reaction solution can be smoothly injected from the PCR sample addition port 105 and the hybridization reagent port 106, making the operation more convenient. Airflow can be pushed through the thrust hole 24 by an external thrust device. Through the push of the external thrust, the airflow passes through the thrust inlet 107, enabling the reaction solution in the PCR reaction chamber 11 to enter the hybridization reaction chamber 12.

[0045] The above - mentioned membrane pump structure has the advantages of being simple and convenient to manufacture. Without connecting other power equipment externally, an external suction device (such as a cylinder - piston suction cylinder) is used to directly achieve automatic control of the first opening - closing component 14, the second opening - closing component 15, the third opening - closing component 16, and the fourth opening - closing component 17 through the four pressure holes 23. It has a high integration degree and high space utilization rate, and is particularly advantageous for use in scenarios where the space is narrow due to equipment limitations and it is impossible to set up external power.

[0046] Please refer to Figure 2 and Figure 3 For reference, in the exemplary embodiment, a discharge pipe 18 is provided at the position of the base 1 corresponding to the discharge port 13. One end of the discharge pipe 18 is connected to the discharge port 13, and the other end is connected to the outside.

[0047] Specifically, one end of the discharge pipe 18 is connected to the discharge port 13, which can enable the waste liquid generated in the hybridization reaction chamber 12 to be smoothly discharged to the outside through the discharge pipe 18.

[0048] Please refer to Figure 2 , in an exemplary embodiment, the PCR reaction cartridge device further includes a PC film 3. The PC film 3 is located between the top cover 2 and the base 1. Through holes are respectively provided at the positions of the PC film 3 corresponding to the PCR sample addition port 105, the hybridization reagent port 106, and the thrust inlet 107.

[0049] Furthermore, pressure-sensitive adhesive is coated at the contact positions of the surface of the PC film 3 with the top cover 2 and the base 1.

[0050] Through holes are respectively provided at the positions of the PC film 3 corresponding to the PCR sample addition port 105, the hybridization reagent port 106, and the thrust inlet 107 to provide channels for sample addition, reagent addition, and solution ejection during the reaction process, while maintaining the sealing of the reaction chamber and the stability of the experimental conditions, ensuring the successful progress of the reaction and the reliability of the results.

[0051] Specifically, pressure-sensitive adhesive (PSA) is coated at the contact positions of the surface of the PC film 3 with the top cover 2 and the base 1. This adhesive can firmly adhere to the surfaces of the top cover 2 and the base 1 after applying appropriate pressure, bonding the base 1 and the top cover 2 together. In this way, the PCR reaction chamber 11 and the hybridization reaction chamber 12 can be effectively sealed to prevent the reaction solution from leaking or being contaminated by external pollutants such as dust and microorganisms from entering the PCR reaction chamber 11 and the hybridization reaction chamber 12, thus avoiding sample contamination, providing good experimental stability, and ensuring the successful progress of the experiment and the reliability of the results.

[0052] It should be noted that the membrane pump structure mentioned in the above embodiment is formed at the positions of the PC film 3 corresponding to the first opening and closing assembly 14, the second opening and closing assembly 15, the third opening and closing assembly 16, and the fourth opening and closing assembly 17, without the need to additionally configure the first opening and closing assembly 14, the second opening and closing assembly 15, the third opening and closing assembly 16, and the fourth opening and closing assembly 17, improving the space utilization rate.

[0053] In an embodiment, an observation port 25 is provided at the position of the top cover 2 corresponding to the hybridization reaction chamber 12, and a transparent plate is fixedly installed at the observation port 25.

[0054] Specifically, the transparent plate can be made of common transparent materials (for example: transparent plastic, transparent glass, etc.). The observation port 25 is provided to facilitate the user to observe the mixing situation and reaction results of the experimental samples during the reaction process. Through the observation port 25, the user can directly check the mixture condition in the hybridization reaction chamber 12, thereby evaluating whether the reaction is proceeding normally or whether the reaction conditions need to be adjusted. It helps to monitor the experimental progress in real time and ensure the accuracy and efficiency of the reaction.

[0055] In an embodiment, a membrane chip (not shown in the figure) is provided in the hybridization reaction chamber 12.

[0056] Specifically, the membrane chip plays a role in separating and immobilizing nucleic acid molecules in the hybridization reaction chamber 12. It can adsorb nucleic acid molecules (such as DNA or RNA), enabling them to stably exist in the reaction system, effectively immobilize the target nucleic acid, avoid its movement and diffusion during the reaction, and is conducive to the subsequent hybridization reaction and result analysis. The membrane chip enables the nucleic acid to effectively hybridize and bind with complementary probes or primers. This immobilization can improve the efficiency and accuracy of the reaction, ensure the specific binding between the target nucleic acid and its complementary probe or primer, thereby enhancing the sensitivity and specificity of the experiment.

[0057] Please refer to Figure 3 , in an exemplary embodiment, the inner bottom surface of the hybridization reaction chamber 12 is an inclined surface.

[0058] Specifically, the ground of the hybridization reaction chamber 12 is set as an inclined surface to facilitate the smooth discharge of the waste liquid generated in the hybridization reaction chamber 12.

[0059] The above are only the preferred embodiments of the present application, and do not limit the patent scope of the present application. Any equivalent structure or equivalent process transformation made by using the content of the specification and drawings of the present application, or directly or indirectly applied in other related technical fields, shall be equally included in the patent protection scope of the present application.

Claims

1. A PCR reaction cartridge device, characterized in that, Comprising: A PCR reaction chamber (11); A hybridization reaction chamber (12), which is communicated with the PCR reaction chamber (11), and a discharge port (13) communicating with the outside is provided at the center of the bottom surface of the hybridization reaction chamber (12); A first flow channel (101), one end of which is communicated with the PCR reaction chamber (11) and the other end of which is communicated with the hybridization reaction chamber (12), and a first opening and closing component (14) is provided on the first flow channel (101); A second flow channel (102), one end of which is communicated with the hybridization reaction chamber (12), and a second opening and closing component (15) is provided on the second flow channel (102); A third flow channel (103), one end of which is communicated with the PCR reaction chamber (11), and a third opening and closing component (16) is provided on the third flow channel (103); A fourth flow channel (104), one end of which is communicated with the third flow channel (103) between the third opening and closing component (16) and the PCR reaction chamber (11), and a fourth opening and closing component (17) is provided on the fourth flow channel (104); A PCR sample adding port (105), which is communicated with the other end of the fourth flow channel (104); A hybridization reagent port (106), which is communicated with the other end of the second flow channel (102); A thrust inlet (107), which is communicated with the other end of the third flow channel (103).

2. The PCR reaction cassette device according to claim 1, wherein Further comprising: A top cover (2); A base (1), a groove is provided on the surface of the base (1) close to the top cover (2), and the base (1) is bonded to the top cover (2) so that the groove forms the PCR reaction chamber (11), the hybridization reaction chamber (12), and the first flow channel (101), the second flow channel (102), the third flow channel (103) and the fourth flow channel (104).

3. The PCR reaction cartridge device according to claim 1, wherein The first opening and closing component (14), the second opening and closing component (15), the third opening and closing component (16), and the fourth opening and closing component (17) all adopt a membrane pump structure.

4. The PCR reaction cartridge device according to claim 2, wherein The top cover (2) includes: A PCR sample adding tube (21), which is arranged at the position of the top cover (2) corresponding to the PCR sample adding port (105), and one end of the PCR sample adding tube (21) penetrates through the top cover (2) and is communicated with the PCR sample adding port (105), and the other end is communicated with the outside; A hybridization reagent tube (22), which is arranged at the position of the top cover (2) corresponding to the hybridization reagent port (106), and one end of the hybridization reagent tube (22) penetrates through the top cover (2) and is communicated with the hybridization reagent port (106), and the other end is communicated with the outside; Four pressure holes (23), which are respectively opened at the positions of the top cover (2) corresponding to the first opening and closing component (14), the second opening and closing component (15), the third opening and closing component (16), and the fourth opening and closing component (17); A thrust hole (24), which is opened at the position of the top cover (2) corresponding to the thrust inlet (107).

5. The PCR reaction cartridge device according to claim 2, characterized in that, A discharge pipe (18) is provided at the position of the base (1) corresponding to the discharge port (13), one end of the discharge pipe (18) is communicated with the discharge port (13), and the other end is communicated with the outside.

6. The PCR reaction cartridge device according to claim 2, wherein Further comprising: A PC film (3) is located between the top cover (2) and the base (1). Through holes are respectively provided at positions of the PC film (3) corresponding to the PCR sample addition port (105), the hybridization reagent port (106), and the thrust inlet (107).

7. The PCR reaction cartridge device according to claim 6, wherein, Pressure-sensitive adhesives are covered at the contact positions of the surface of the PC film (3) with the top cover (2) and the base (1).

8. The PCR reaction cartridge device according to claim 2, wherein An observation port (25) is provided at a position of the top cover (2) corresponding to the hybridization reaction chamber (12), and a transparent plate is fixedly installed at the observation port (25).

9. The PCR reaction cartridge device according to claim 8, wherein A membrane chip is provided in the hybridization reaction chamber (12).

10. The PCR reaction cartridge device according to claim 9, wherein, The inner bottom surface of the hybridization reaction chamber (12) is an inclined surface.

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