Multi-channel nucleic acid reaction chip, gene detection device and gene detection instrument

Abstract

The utility model discloses a multi-channel nucleic acid reaction chip, a gene detection device and a gene detection instrument, relates to the technical field of molecular diagnosis, and aims to improve the condition that liquid crosstalk occurs among reaction cavities in a nucleic acid amplification reaction process to influence a nucleic acid detection result. The multi-channel nucleic acid reaction chip is used for being connected with a gene detection kit and comprises a first flow channel, a plurality of mutually independent reaction channels and an exhaust channel. One end of the first flow channel is used for being communicated with a liquid outlet of a gene detection kit; one end of each reaction channel is communicated with the first flow channel, each reaction channel comprises a nucleic acid amplification reaction cavity and a branch valve body, and the nucleic acid amplification reaction cavity is communicated with the branch valve body; the exhaust passage comprises an exhaust valve body, and the exhaust valve body communicates with the other end of the first flow channel. Therefore, the method has the advantage of improving the accuracy and reliability of nucleic acid detection.

Description

Technical Field

[0001] The present application relates to the technical field of molecular diagnosis, and in particular to a multi-channel nucleic acid reaction chip, a gene detection device and a gene detection instrument. Background Art

[0002] In the field of molecular diagnostics, multi-channel nucleic acid detection and analysis platforms for multiple indicators in a single sample have become an important research direction. Multi-channel nucleic acid reaction chips, one such platform for rapid and efficient nucleic acid detection, can miniaturize and integrate traditional molecular biology experimental processes. This technology allows for simultaneous detection of multiple target sequences within the same sample on a compact chip.

[0003] The nucleic acid reaction chip can adopt a method of setting up multiple reaction flow paths separately, and inject the sample solution into each reaction flow path separately to perform the nucleic acid reaction. However, this method requires corresponding instruments or manual alignment and injection of each reaction flow path one by one. Its operation is complicated, the structure is cumbersome, and it significantly reduces the efficiency of nucleic acid detection. The nucleic acid reaction chip can also adopt a one-way multiple flow path design, injecting the sample solution in the main path into each branch path, and then allowing the sample solution to flow into the reaction chamber set in each branch path for nucleic acid reaction. However, since the temperature needs to be increased or the temperature cycle is required during the nucleic acid reaction process, the sample solution in the reaction cavity and the branch path can easily diffuse to other areas through the connection position between the branch path and the main path due to temperature changes, causing liquid crosstalk, thereby affecting the accuracy of the nucleic acid detection results.

[0004] Some chips use hot-melt paraffin to seal the flow path in the front section of the reaction chamber, closing off each branch to prevent liquid crosstalk. However, the reliability of this seal is difficult to guarantee. Due to factors such as the density and brittleness of paraffin, the chip still faces the risk of crosstalk between the sample solution and the melted paraffin contaminating the solution in the reaction chamber. In addition, paraffin may cause optical interference with fluorescence detection during nucleic acid reactions, resulting in significant limitations.

[0005] Therefore, how to design a multi-channel nucleic acid reaction chip that can realize single-sample multi-index nucleic acid detection while effectively reducing solution diffusion crosstalk caused by temperature changes, thereby improving the accuracy and reliability of nucleic acid detection, has become a topic that urgently needs research in the field of molecular diagnosis. Utility Model Content

[0006] The purpose of this application is to provide a multi-channel nucleic acid reaction chip, a gene detection device and a gene detection instrument, which can reduce the probability of liquid crosstalk between reaction chambers due to temperature increase during the nucleic acid amplification reaction affecting the detection results, and improve the accuracy and reliability of nucleic acid detection.

[0007] The embodiment of the present application is implemented as follows:

[0008] In a first aspect, embodiments of the present application provide a multi-channel nucleic acid reaction chip for connection to a genetic testing kit, the multi-channel nucleic acid reaction chip comprising a first flow channel, a plurality of independent reaction pathways, and an exhaust pathway. One end of the first flow channel is connected to the liquid outlet of the genetic testing kit; one end of each reaction pathway is connected to the first flow channel; the reaction pathway comprises a nucleic acid amplification reaction chamber and a branch valve body, the nucleic acid amplification reaction chamber and the branch valve body being connected; and the exhaust pathway comprises an exhaust valve body, which is connected to the other end of the first flow channel.

[0009] In the above technical solution, the multi-channel nucleic acid reaction chip realizes the injection of the same sample solution into multiple reaction chambers through a one-way, multi-pathway structure design; by setting a branch valve body in each reaction channel, the fluid flow in each reaction channel can be controlled; by connecting the exhaust channel provided with an exhaust valve body at the other end of the first flow channel, after each nucleic acid amplification reaction chamber is filled with sample solution, gas can be injected into the exhaust channel through the first flow channel to achieve air isolation at the connection position between each reaction channel and the first flow channel, so as to reduce the probability that the sample solution in the nucleic acid amplification reaction chamber flows and diffuses to other nucleic acid amplification reaction chambers due to temperature changes, causing liquid crosstalk and affecting the detection results, thereby improving the accuracy and reliability of the nucleic acid detection results.

[0010] In some embodiments, the reaction pathway further includes at least one branch liquid detection point, which is connected to the nucleic acid amplification reaction chamber via a branch flow channel, or alternatively, the branch liquid detection point is located within the nucleic acid amplification reaction chamber. In the above technical solution, the provision of branch liquid detection points in each reaction pathway can control the amount of sample solution injected into the corresponding nucleic acid amplification reaction chamber, thereby reducing sample solution waste and improving the uniformity of the sample solution within each reaction chamber.

[0011] In some embodiments, the exhaust pathway further includes at least one exhaust detection point, which is connected to the exhaust valve body via the exhaust flow channel. In the above technical solution, the provision of exhaust detection points within the exhaust pathway enables detection of whether the sample solution at the connection between each reaction channel and the first flow channel has been displaced and emptied by gas, thereby improving the reliability of gas isolation between the reaction channels.

[0012] In some embodiments, the connection point between each reaction pathway and the first flow channel is a branch connection point, and the end of the first flow channel that is connected to the liquid outlet is the liquid inlet end. The multi-channel nucleic acid reaction chip also includes a pre-liquid detection point, which is located in the first flow channel and is located between the liquid inlet end and the branch connection point closest to the liquid inlet end. In the above technical solution, the provision of the pre-liquid detection point can ensure that the airtightness of each pathway in the chip is tested when the sample solution flows between the piston chamber and the amplification enzyme ball chamber of the genetic testing kit, thereby improving the reliability of the directional flow of the sample solution.

[0013] In some embodiments, the connection point between each reaction pathway and the first flow channel is a branch connection point, and the end of the first flow channel that is connected to the liquid outlet is a liquid inlet end. The multi-channel nucleic acid reaction chip also includes a pre-valve body, which is disposed in the first flow channel and is located between the liquid inlet end and the branch connection point closest to the liquid inlet end. In the above technical solution, the provision of the pre-valve body can improve the reliability of the sample solution flowing or directional flow within a designated area of ​​the multi-channel nucleic acid reaction chip.

[0014] In some embodiments, the genetic testing kit includes a first liquid outlet and a second liquid outlet, and the multi-channel nucleic acid reaction chip further includes a first branch and a second branch; the first branch and the second branch are connected at the liquid inlet end of the first flow channel, the first branch is used to communicate with the first liquid outlet, and the second branch is used to communicate with the second liquid outlet. In the above technical solution, the provision of the first branch and the second branch enables the sample solution to flow between the two cavities of the genetic testing kit, thereby improving the practicality of the multi-channel nucleic acid reaction chip.

[0015] In some embodiments, the valve body in the multi-channel nucleic acid reaction chip includes an input through hole, an output through hole and a stop valve cavity, one end of the input through hole and one end of the output through hole are connected to the stop valve cavity, and the other end of the input through hole and the other end of the output through hole are connected to the flow channel; the stop valve cavity can seal one end of the input through hole and / or one end of the output through hole by squeezing the valve core column.

[0016] In some embodiments, multiple valve bodies in the multi-channel nucleic acid reaction chip are distributed on the same straight line. In the above technical solution, multiple valve bodies are distributed on the same straight line, so that the corresponding valve control module can achieve simplified structure and control logic.

[0017] In some embodiments, the multi-channel nucleic acid reaction chip further includes multiple discharge chambers that communicate with the reaction pathways and / or the exhaust pathways. In this technical solution, the provision of discharge chambers further simplifies the overall structure of the genetic testing device, eliminating the need for separate discharge piping and waste gas and liquid collection boxes; waste gas and liquid collection can be achieved solely through the multi-channel nucleic acid reaction chip.

[0018] In some embodiments, the multi-channel nucleic acid reaction chip further includes at least one partitioning space disposed between two adjacent nucleic acid amplification reaction chambers. In the above technical solution, the provision of the partitioning space can reduce fluorescence crosstalk between adjacent nucleic acid amplification reaction chambers, thereby improving the accuracy and reliability of nucleic acid detection.

[0019] In some embodiments, the centers of the multiple nucleic acid amplification reaction chambers are located on the same straight line. The above technical solution makes the structural layout of the multi-channel nucleic acid reaction chip more reasonable and can also make the overall structure of the corresponding fluorescence acquisition and analysis module more compact.

[0020] In some embodiments, the nucleic acid amplification reaction chamber includes a reaction inlet and a reaction outlet; when the multi-channel nucleic acid reaction chip is inserted into the genetic testing kit in an upright position, the reaction inlet is located at the bottom of the nucleic acid amplification reaction chamber, and the reaction outlet is located at the top of the nucleic acid amplification reaction chamber. In the above technical solution, the reaction inlet is located at the bottom of the nucleic acid amplification reaction chamber, and the reaction outlet is located at the top of the nucleic acid amplification reaction chamber. This makes it easier for the nucleic acid amplification reaction chamber to be filled with the sample solution, reduces the probability of bubbles remaining in the reaction chamber, and thereby improves the uniformity of the sample solution in each reaction chamber.

[0021] In some embodiments, a multi-channel nucleic acid reaction chip includes a substrate and a sealing film. The substrate has a first surface and a second surface disposed opposite each other; grooves and / or through-holes are formed on the first surface and / or the second surface. The sealing film covers the first surface and the second surface, respectively, and cooperates with the substrate to form a first flow channel, a nucleic acid amplification reaction chamber, a branch valve body, and an exhaust valve body. In the above technical solution, the method of forming grooves on the substrate and cooperating with the sealing film to form the corresponding flow channel, valve body, and reaction chamber reduces the processing difficulty and manufacturing cost of the multi-channel nucleic acid reaction chip and facilitates observation.

[0022] In some embodiments, a multi-channel nucleic acid reaction chip includes a chip body and a plug-in interface, wherein a first flow channel is disposed in the chip body; the plug-in interface is in communication with the first flow channel and connected to the chip body, and is configured to interface with the liquid outlet. In the above technical solution, the provision of the plug-in interface allows for a more reliable connection between the multi-channel nucleic acid reaction chip and the genetic testing kit at the liquid outlet, reducing the probability of leakage and aerosol diffusion contamination.

[0023] In the second aspect, an embodiment of the present application provides a gene detection device, which includes a multi-channel nucleic acid reaction chip, a valve control module, a fluorescence collection and analysis module, and a gene detection kit provided by any embodiment of the first aspect of the present application. Among them, the gene detection kit has a liquid outlet; one end of the multi-channel nucleic acid reaction chip is connected to the gene detection kit at the liquid outlet; the valve control module is arranged on one side of the multi-channel nucleic acid reaction chip, and is adjacent to each valve body of the multi-channel nucleic acid reaction chip; the fluorescence collection and analysis module has a fluorescence collection hole, and the multi-channel nucleic acid reaction chip has one end of a nucleic acid amplification reaction chamber, which extends into and is accommodated in the fluorescence collection hole. In the above technical solution, the gene detection device has the same technical effect as the aforementioned multi-channel nucleic acid reaction chip.

[0024] In some embodiments, when multiple valve bodies in a multi-channel nucleic acid reaction chip are distributed on the same straight line, the valve control module includes a mounting seat, a camshaft, a drive motor, and multiple valve core columns. The camshaft is rotatably connected to the mounting seat; the output shaft of the drive motor is connected to the camshaft; multiple valve core columns are arranged along the same straight line and accommodated in the various valve core column holes of the mounting seat, and the camshaft includes multiple protrusions; one end of the valve core column rests on the protrusion, and the other end of the valve core column is aligned with the valve body. In the above technical solution, the cooperation between the various protrusions on the camshaft and the valve core column enables the valve control module to achieve on-off control of different valve bodies through a single power source, and the structure and control of the valve control module are significantly simplified.

[0025] In a third aspect, an embodiment of the present application provides a genetic testing instrument, which includes the genetic testing device provided by any embodiment of the second aspect of the present application. In the above technical solution, the genetic testing instrument has the same technical effects as the aforementioned genetic testing device. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] In order to more clearly illustrate the technical solutions of the embodiments of the present application, the following is a brief introduction to the drawings required for use in the embodiments. It should be understood that the following drawings only show certain embodiments of the present application and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other relevant drawings can be obtained based on these drawings without creative work.

[0027] Figure 1 This is a schematic diagram of the structure of a genetic testing instrument according to one embodiment of the present application;

[0028] Figure 2 This is a schematic diagram of the overall structure of a gene detection device according to one embodiment of the present application;

[0029] Figure 3 This is a disassembled schematic diagram of a gene detection device according to one embodiment of the present application;

[0030] Figure 4 This is a schematic structural diagram of a gene detection kit and a multi-channel nucleic acid reaction chip in a docking state according to an embodiment of the present application;

[0031] Figure 5 This is a cross-sectional schematic diagram of a gene detection kit and a multi-channel nucleic acid reaction chip in a docking state according to an embodiment of the present application;

[0032] Figure 6 This is a schematic structural diagram of the first side of a multi-channel nucleic acid reaction chip shown in the first embodiment of the present application;

[0033] Figure 7 This is a schematic diagram of the manufacturing process of a multi-channel nucleic acid reaction chip according to one embodiment of the present application;

[0034] Figure 8 This is a schematic structural diagram of the first side of a multi-channel nucleic acid reaction chip according to the second embodiment of the present application;

[0035] Figure 9 This is a schematic structural diagram of the first side of a multi-channel nucleic acid reaction chip according to the third embodiment of the present application;

[0036] Figure 10 This is a schematic structural diagram of the first side of a multi-channel nucleic acid reaction chip according to the fourth embodiment of the present application;

[0037] Figure 11 This is a schematic diagram of the second side structure of the multi-channel nucleic acid reaction chip shown in the second embodiment of the present application;

[0038] Figure 12 This is a schematic diagram of the valve body structure shown in one embodiment of the present application;

[0039] Figure 13 This is a schematic diagram showing a valve body in a closed state according to an embodiment of the present application;

[0040] Figure 14 This is a schematic diagram of the overall structure of a valve control module according to an embodiment of the present application;

[0041] Figure 15 This is an exploded schematic diagram of a valve control module according to an embodiment of the present application;

[0042] Figure 16 This is a schematic diagram of a partial structure of a valve control module according to an embodiment of the present application;

[0043] Figure 17 A schematic cross-sectional view of a valve control module according to an embodiment of the present application;

[0044] Figure 18 This is a schematic structural diagram of a fluorescence collection and analysis module according to an embodiment of the present application;

[0045] Figure 19 This is a schematic diagram of a multi-channel nucleic acid reaction chip loaded with a fluorescence acquisition and analysis module according to one embodiment of the present application.

[0046] Icons: 1-Gene detection instrument; 2-Gene detection device; 3-Gene detection kit; 4-Multi-channel nucleic acid reaction chip; 5-Valve control module; 6-Fluorescence acquisition and analysis module; 30-Screw cap; 31-Piston chamber; 32-Amplification enzyme ball chamber; 300-Liquid outlet; 301-First liquid outlet; 302-Second liquid outlet; 40-Chip body; 401-Substrate; 402-Sealing film; 41-Input channel; 4101-Liquid inlet end; 4102-Branch connection position; 411-First branch; 412-Second branch; 413-First flow channel; 414-Pre-valve body; 42-Reaction channel; 4201-First reaction channel; 4202-Second reaction channel; 4203-Third Reaction pathway; 4204-fourth reaction pathway; 421-nucleic acid amplification reaction chamber; 4211-reaction inlet; 4212-reaction outlet; 422-branch flow channel; 423-branch valve body; 424-branch liquid detection point; 43-exhaust pathway; 431-exhaust valve body; 432-exhaust flow channel; 44-valve body; 441-input through hole; 442-output through hole; 440-stop valve chamber; 45-discharge chamber; 46-partition space; 47-plug-in interface; 50-mounting seat; 501-valve core column hole; 51-drive motor; 52-coupling; 53-camshaft; 531-protrusion; 54-valve core column; 55-elastic member; 60-fluorescence collection hole; 61-fixed bracket. DETAILED DESCRIPTION

[0047] The terms "first", "second", "third", etc. are only used to distinguish and describe, and do not indicate the order of arrangement, nor can they be understood as indicating or implying relative importance.

[0048] Furthermore, terms such as "horizontal," "vertical," and "overhanging" do not necessarily imply that a component must be absolutely horizontal or overhanging, but rather that it can be slightly tilted. For example, "horizontal" simply means that its direction is more horizontal than "vertical," and does not mean that the structure must be completely horizontal, but rather that it can be slightly tilted.

[0049] In the description of this application, it should be noted that the terms "inside", "outside", "left", "right", "up", "down", etc. indicate directions or positional relationships based on the directions or positional relationships shown in the accompanying drawings, or are the directions or positional relationships in which the product of the application is usually placed when 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 direction, be constructed and operated in a specific direction. Therefore, they should not be understood as limitations on this application.

[0050] In the description of this application, unless otherwise clearly specified and limited, the terms "set", "install", "connected" and "connect" should be understood in a broad sense. For example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection or an indirect connection through an intermediate medium, or it can be a connection between the internal parts of two elements.

[0051] The technical solution of this application will be described in detail below with reference to the accompanying drawings.

[0052] Molecular diagnostic technology based on the principle of PCR (Polymerase Chain Reaction) amplification is a technique that detects the presence or absence of endogenous target genes (genetic or variant) or exogenous target genes (pathogens) by specifically amplifying target genes through primers, so as to provide key information and decision-making basis for disease diagnosis and treatment methods. Its main application scenarios include infectious disease diagnosis, blood screening, tumor mutation site detection, genetic disease diagnosis, prenatal diagnosis, tissue typing, etc. Molecular diagnostic technology based on PCR amplification generally includes the following steps: sample lysis, nucleic acid purification, nucleic acid amplification under the constraints of specific primers, and fluorescence signal collection and analysis.

[0053] In molecular diagnostic systems based on PCR amplification, due to the possibility of aerosol contamination during PCR amplification and to avoid cross-contamination between samples, it is usually necessary to set up a partitioned laboratory for sample processing, nucleic acid extraction, PCR amplification and other partitioned operations. This partitioned laboratory must have a good ventilation system, but the construction cost is relatively high. In addition, the configuration of experimental operators will also increase labor costs. Excessive manual intervention can easily increase the probability of experimental errors and incorrect test results. The aforementioned problems will increase the technical threshold for the use of molecular diagnostic platforms manufactured based on the principle of PCR amplification.

[0054] On this basis, the process quality control requirements for molecules to complete multi-sample and multi-test project experiments are very high, and molecular diagnostic laboratories are suitable for multi-sample single-indicator detection modes. It is difficult to efficiently adapt to single-sample multi-indicator detection modes, and its detection indicators are limited. For example, it is difficult to achieve single-sample multi-indicator infection pathogen screening. With the current clinical diagnosis needs, molecular diagnosis has been moving towards the direction of being able to detect all pathogens that may cause the disease at one time, so as to analyze the exact cause of the disease at one time. For example, in the etiology diagnosis of respiratory diseases or sexually transmitted diseases, designing a detection platform that can detect more target genes has become a key trend in product research and development in the field of molecular diagnosis. As a result, multi-channel nucleic acid reaction chips that can realize multiple PCR amplification detection came into being.

[0055] The multi-channel nucleic acid reaction chip is an integrated microfluidic container, which is usually used for efficient and rapid nucleic acid (DNA or RNA) detection and analysis. This chip can be used independently or in conjunction with a small genetic detection kit, integrating traditional laboratory operation steps (such as sample lysis, elution, purification or nucleic acid amplification) on a miniaturized platform, realizing automated and portable nucleic acid detection and analysis. In the related art, a multi-channel nucleic acid reaction chip is provided with a plurality of microfluidic channels connected to the same input flow channel for sample introduction, mixing, reaction, etc. However, during the nucleic acid amplification reaction process, due to temperature increase or temperature cycling, liquid crosstalk is likely to occur between the various microfluidic channels on the multi-channel nucleic acid reaction chip, thereby affecting the detection results.

[0056] Therefore, how to design a multi-channel nucleic acid reaction chip that can realize single-sample multi-index nucleic acid detection while effectively reducing solution diffusion crosstalk caused by temperature changes, thereby improving the accuracy and reliability of nucleic acid amplification detection, has become a key research direction in the design of nucleic acid reaction chips in the field of molecular diagnosis.

[0057] Based on the above considerations, an embodiment of the present application provides a multi-channel nucleic acid reaction chip, which realizes the injection of the same sample solution from the same input flow channel into multiple reaction chambers (or multiple reaction channels, multiple microfluidic channels) through a one-through-multiple channel structure design; by setting a branch valve body in each reaction channel, the fluid flow in each reaction channel is controlled; by connecting an exhaust channel with an exhaust valve body at the end of the input flow channel, the chip can inject gas into the exhaust channel through the input flow channel after each nucleic acid amplification reaction chamber is filled with sample solution, so as to achieve air isolation at the connection position between each reaction channel and the input flow channel, thereby reducing the probability that the sample solution in the nucleic acid amplification reaction chamber flows and diffuses into other nucleic acid amplification reaction chambers due to temperature changes, causing liquid crosstalk and affecting the detection results, thereby improving the accuracy of the nucleic acid detection results and the reliability of nucleic acid detection.

[0058] See Figure 1 , Figure 1 This is a schematic diagram of the structure of a gene detection instrument 1 according to an embodiment of the present application. Figure 1 As shown, the genetic detection instrument 1 in the embodiment of the present application may include at least one genetic detection device 2, which refers to a device that can process biological samples, amplify target nucleic acid fragments in the sample solution through PCR technology, and quantitatively analyze nucleic acids by real-time monitoring of fluorescence signals.

[0059] In some embodiments, the genetic testing instrument 1 may further include a controller (or control system), which may be electrically connected to the various components in the genetic testing device 2 to issue control signals to control the genetic testing device 2 to automatically perform corresponding experimental operations at each stage.

[0060] See Figures 2 to 3 , Figure 2 This is a schematic structural diagram of a gene detection device 2 according to an embodiment of the present application; Figure 3 This is a disassembled schematic diagram of a gene detection device 2 according to an embodiment of the present application. Figures 2 to 3 As shown, the gene detection device 2 in the embodiment of the present application may include a multi-channel nucleic acid reaction chip 4, a valve control module 5, a fluorescence collection and analysis module 6 and a gene detection kit 3.

[0061] Among them, the gene detection kit 3 can be connected and communicated with the multi-channel nucleic acid reaction chip 4; the valve control module 5 can be arranged on one side of the multi-channel nucleic acid reaction chip 4, and arranged adjacent to each valve body 44 in the multi-channel nucleic acid reaction chip 4; the fluorescence collection and analysis module 6 can have a fluorescence collection hole 60, and the multi-channel nucleic acid reaction chip 4 has one end of the nucleic acid amplification reaction chamber 421, which can be extended into and accommodated in the fluorescence collection hole 60.

[0062] In the embodiment of the present application, the gene detection kit 3 refers to a container that can perform preliminary processing such as sample lysis, nucleic acid elution, and nucleic acid purification on biological samples; the multi-channel nucleic acid reaction chip 4 refers to a closed container that can accommodate the processed sample solution and amplify it at a specified position under the constraint of specific primers to produce a fluorescent reaction; the fluorescence acquisition and analysis module 6 refers to a module that can collect signals from fluorescent groups and perform strength analysis based on the collected fluorescence signals to determine whether a specific DNA / RNA target sequence exists in the biological sample; the valve control module 5 refers to a module that can control the on and off of each valve body 44 in the multi-channel nucleic acid reaction chip 4, and thereby control the flow direction and on and off of the sample solution in each flow channel or passage.

[0063] See Figures 4 to 6 , Figure 4 This is a structural diagram of a gene detection kit 3 and a multi-channel nucleic acid reaction chip 4 in a docking state according to an embodiment of the present application; Figure 5 This is a cross-sectional schematic diagram of a gene detection kit 3 and a multi-channel nucleic acid reaction chip 4 in a docking state according to an embodiment of the present application; Figure 6 This is a schematic diagram of the first side structure of the multi-channel nucleic acid reaction chip 4 shown in the first embodiment of the present application. Figures 4 to 6 As shown, the gene detection kit 3 has at least one liquid outlet 300 ; one end (ie, the input path 41 ) of the multi-channel nucleic acid reaction chip 4 can be docked with and communicated with the gene detection kit 3 at the liquid outlet 300 .

[0064] In the embodiment of the present application, the multi-channel nucleic acid reaction chip 4 is used to connect to the gene detection kit 3. The multi-channel nucleic acid reaction chip 4 may include multiple independent reaction pathways 42 and exhaust pathways 43. The input pathway 41 includes a first flow channel 413, one end of which can be used to communicate with the liquid outlet 300 of the gene detection kit 3; one end of each reaction pathway 42 can be connected to the first flow channel 413, and the reaction pathway 42 includes a nucleic acid amplification reaction chamber 421 and a branch valve body 423, and the nucleic acid amplification reaction chamber 421 is connected to the branch valve body 423; the exhaust pathway 43 includes an exhaust valve body 431, and the exhaust valve body 431 is connected to the other end of the first flow channel 413.

[0065] In the above technical solution, the multi-channel nucleic acid reaction chip 4 realizes the injection of the same sample solution into multiple reaction chambers through a one-way multi-channel structure design; by setting a branch valve body 423 in each reaction channel 42, the fluid flow in each reaction channel 42 can be controlled; by connecting the exhaust channel 43 provided with an exhaust valve body 431 at the other end of the first flow channel 413, the chip can inject gas into the exhaust channel 43 through the first flow channel after each nucleic acid amplification reaction chamber 421 is filled with sample solution, thereby isolating the air at the connection position between each reaction channel 42 and the first flow channel, thereby reducing the probability that the sample solution in the nucleic acid amplification reaction chamber 421 flows and diffuses into other nucleic acid amplification reaction chambers 421 due to temperature changes, causing liquid crosstalk and affecting the detection results, thereby improving the accuracy and reliability of the nucleic acid detection results.

[0066] like Figure 6 As shown, in some embodiments, the multi-channel nucleic acid reaction chip 4 may include a chip body 40 and at least one plug-in interface 47. The nucleic acid amplification reaction chamber 421, the branch valve body 423, the exhaust valve body 431, and the first flow channel 413 are all disposed in the chip body 40; the plug-in interface 47 is disposed at one end of the chip body 40. The plug-in interface 47 can communicate with the first flow channel 413 and connect to the chip body 40. The plug-in interface 47 is used to dock with the liquid outlet 300. In the above technical solution, the provision of the plug-in interface 47 makes the connection between the multi-channel nucleic acid reaction chip 4 and the genetic testing kit 3 at the liquid outlet 300 more reliable, reduces the probability of leakage, and reduces the diffusion and contamination of aerosols.

[0067] See Figure 7 , Figure 7 FIG. 4 is a schematic diagram of a manufacturing process of a multi-channel nucleic acid reaction chip 4 according to an embodiment of the present application. Figure 7As shown, the multi-channel nucleic acid reaction chip 4 may include a substrate 401 and two sealing films 402. The substrate 401 has a first surface and a second surface disposed opposite to each other. A plurality of grooves, through-holes, concave surfaces, and other structures are formed on the first surface and / or the second surface. The sealing films 402 may be welded, glued, or bonded to cover the first surface and the second surface, respectively, to cooperate with the grooves, through-holes, and concave surfaces on the substrate 401 to form structures such as the first flow channel 413, the nucleic acid amplification reaction chamber 421, the branch valve body 423, and the exhaust valve body 431.

[0068] Coating both the front and back surfaces of substrate 401 allows the grooves, through-holes, and concave surfaces of chip body 40 to form sealed chambers, which facilitate the flow, storage, and reaction of sample solutions within chip body 40. In the above technical solution, the formation of grooves on substrate 401, which cooperate with sealing film 402 to form corresponding flow channels, valve body 44, and chambers, simplifies the structure of multi-channel nucleic acid reaction chip 4, reduces processing difficulty and manufacturing costs, and facilitates observation when the sealing film is transparent.

[0069] Please combine Figures 4 to 6 As shown, in some embodiments, the gene detection kit 3 includes a first liquid outlet 301 and a second liquid outlet 302. Corresponding to the first liquid outlet 301 and the second liquid outlet 302, the input path 41 of the multi-channel nucleic acid reaction chip 4 may further include a first branch 411 and a second branch 412; the first branch 411 and the second branch 412 may be connected at the liquid inlet end 4101 of the first flow channel 413. The first branch 411 may be used to communicate with the first liquid outlet 301, and the second branch 412 may be used to communicate with the second liquid outlet 302. Specifically, one end of the first branch 411 is connected to the first liquid outlet 301, one end of the second branch 412 is connected to the second liquid outlet 302, and the other end of the first branch 411 and the other end of the second branch 412 are connected at the liquid inlet end 4101 of the first flow channel 413. The liquid inlet end 4101 of the first flow channel 413 refers to the end of the first flow channel 413 that is close to the liquid outlet 300 of the genetic testing kit 3. In the above technical solution, the provision of the first branch 411 and the second branch 412 enables the sample solution to flow between the two chambers of the genetic testing kit 3, thereby improving the practicality of the multi-channel nucleic acid reaction chip 4 and expanding its scope of application.

[0070] like Figure 5As shown, the genetic testing kit 3 may include at least two chambers. One chamber is a piston chamber 31, the bottom end of which can be connected to the first liquid outlet 301, and a piston rod is provided in the piston chamber 31. The piston rod is used to squeeze the sample solution in the piston chamber 31 into other flow channels or chambers, or to draw the sample solution from other chambers or flow channels by stretching to create a negative pressure environment; the other chamber is an amplification enzyme ball chamber 32, the bottom end of which can be connected to the second liquid outlet 302, and a piston rod is also provided in the amplification enzyme ball chamber 32. The amplification enzyme ball chamber 32 can be filled with a spherical PCR amplification enzyme. The purified nucleic acid solution needs to be combined with the PCR amplification enzyme to perform the PCR amplification reaction.

[0071] In one application, the gene detection device 2 can first dissolve the amplification enzyme ball in the amplification enzyme ball cavity 32 into the sample solution by connecting the gene detection kit 3 to the multi-channel nucleic acid reaction chip 4 and setting the first branch 411 and the second branch 412 on the multi-channel nucleic acid reaction chip 4. The specific control operation process is as follows: After the test sample completes the steps of sample lysis, nucleic acid washing, and nucleic acid elution in the gene detection kit 3, the purified nucleic acid solution is pumped into the piston cavity 31 of the gene detection kit 3. The PCR amplification enzyme can be pre-made into spherical particles and stored in an amplification enzyme ball cavity 32 of the kit. At this time, the nucleic acid solution can be first injected into the amplification enzyme ball cavity 32 to melt the amplification enzyme ball in the nucleic acid solution.

[0072] In this step, all valve bodies 44 in the multi-channel nucleic acid reaction chip 4 can be closed in advance, and then the piston rod in the piston chamber 31 is pressed down. Since each reaction channel 42 and exhaust channel 43 in the multi-channel nucleic acid reaction chip 4 are in a blocked state, the nucleic acid solution will enter the first branch 411 through the first liquid outlet 301, then enter the second branch 412 through the liquid inlet end 4101 of the first flow channel 413, and then enter the amplification enzyme ball chamber 32 through the second liquid outlet 302 and melt the amplification enzyme ball. After the primer ball (amplification enzyme ball) is completely melted, the piston rod in the piston chamber 31 is controlled to be pulled upward, and the nucleic acid solution (also referred to as the resolution solution or sample solution in the following embodiment) that has dissolved the PCR amplification enzyme ball can be withdrawn to the piston chamber 31, waiting to be injected into the multi-channel nucleic acid reaction chip 4.

[0073] like Figure 6As shown, in some embodiments, the multi-channel nucleic acid reaction chip 4 further includes a plurality of discharge chambers 45, and the discharge chambers 45 are connected to the reaction pathway 42 and / or the exhaust pathway 43. In the embodiment of the present application, each reaction pathway 42 or exhaust pathway 43 is connected to a discharge chamber 45. The discharge chamber 45 refers to a chamber in the multi-channel nucleic acid reaction chip 4 for storing excess sample solution or gas. In the above technical solution, through the provision of the discharge chamber 45, the overall structure of the gene detection device 2 can be further simplified, and the gene detection device 2 does not need to separately connect the discharge pipeline and the waste gas and waste liquid box to collect excess gas or sample solution, and the waste gas and waste liquid collection can be achieved only through the multi-channel nucleic acid reaction chip 4.

[0074] In addition, the multi-channel nucleic acid reaction chip 4 can also reduce the probability of excessive injection of excess solution and gas into the discharge chamber and subsequent rupture by setting up branch liquid detection points 424, exhaust detection points or pre-liquid detection points, thereby improving the reliability of genetic testing.

[0075] See Figures 8 to 10 , Figure 8 This is a schematic structural diagram of the first side of a multi-channel nucleic acid reaction chip 4 shown in the second embodiment of the present application; Figure 9 This is a schematic structural diagram of the first side of a multi-channel nucleic acid reaction chip 4 shown in the third embodiment of the present application; Figure 10 This is a schematic diagram of the first side structure of the multi-channel nucleic acid reaction chip 4 shown in the fourth embodiment of this application. Figures 5 to 10 As shown, the reaction pathway 42 may further include at least one branch liquid detection point 424. The branch liquid detection point 424 may be connected to the nucleic acid amplification reaction chamber 421 via the branch flow channel 422, or the branch liquid detection point 424 may be directly disposed within the nucleic acid amplification reaction chamber 421. In the above technical solution, by disposing the branch liquid detection point 424 in each reaction pathway 42, the amount of sample solution injected into the corresponding nucleic acid amplification reaction chamber 421 can be controlled, thereby reducing sample solution waste, improving the uniformity of the sample solution within each reaction chamber, and thereby improving the accuracy of the test results.

[0076] The purpose of setting up branch liquid detection points 424 in each reaction pathway 42 is to fill each nucleic acid amplification reaction chamber 421 more effectively and equally. Due to the differences in enzymes, temperature, and volume of injected nucleic acid solution in the reaction system during the PCR reaction, the test results will vary greatly. For example, during the liquid injection process, if the nucleic acid amplification reaction chamber 421 in the first reaction pathway 4201 is completely filled, while the nucleic acid amplification reaction chamber 421 in the third reaction pathway 4203 is only filled to two-thirds, performing a PCR reaction based on this situation will cause the reaction curve of the final test result corresponding to the third reaction pathway 4203 to have a very weak peak or even no peak at all. In this case, the result will be directly judged as negative when the result is determined, but this is actually very likely to be a misjudgment. Therefore, it is of great significance to improve the consistency of the reaction system (reaction parameters) in each reaction chamber.

[0077] like Figure 8 As shown, in some embodiments, the branch liquid detection points 424 in each reaction channel 42 can be located between the reaction outlet 4212 of the nucleic acid amplification reaction chamber 421 and the branch valve body 423; alternatively, the branch valve body 423 can be located in the branch flow channel 422 on the side of the reaction inlet 4211 of the nucleic acid amplification reaction chamber 421, and the branch liquid detection points 424 can be located in the branch flow channel 422 on the side of the reaction outlet 4212 of the nucleic acid amplification reaction chamber 421. During the process of injecting sample solution into the nucleic acid amplification reaction chambers 421 in each reaction channel 42 one by one, as long as liquid is detected at the branch liquid detection points 424 on the side of the reaction outlet 4212 of the nucleic acid amplification reaction chamber 421, it can be determined that the current nucleic acid amplification reaction chamber 421 is filled with sample solution, and the branch valve body 423 in the reaction channel 42 can be further controlled to close to stop injection.

[0078] like Figure 9 As shown, in some embodiments, the branch liquid detection point 424 in each reaction channel 42 can be located in the branch flow channel 422 on the side of the reaction inlet 4211 of the nucleic acid amplification reaction chamber 421, that is, the branch liquid detection point 424 is located in front of the nucleic acid amplification reaction chamber 421. During the process of injecting sample solution into the nucleic acid amplification reaction chamber 421 in each reaction channel 42 one by one, when liquid is detected at the branch liquid detection point 424, a timer can be automatically started. The end of the timer indicates that the sample solution has just filled the nucleic acid amplification reaction chamber 421. The timer duration can be adaptively adjusted according to the chamber capacity. The larger the chamber capacity, the longer the timer duration, and the smaller the chamber capacity, the shorter the timer duration.

[0079] like Figure 10As shown, in some embodiments, the branch liquid detection points 424 in each reaction channel 42 are all located within the nucleic acid amplification reaction chamber 421. Furthermore, the branch liquid detection points 424 can be located at the reaction outlet 4212 or at the topmost portion of the nucleic acid amplification reaction chamber 421. During the process of injecting sample solution into the nucleic acid amplification reaction chamber 421 in each reaction channel 42 one by one, when liquid is detected at the branch liquid detection points 424, it indicates that the sample solution has just filled the nucleic acid amplification reaction chamber 421.

[0080] In some embodiments, the exhaust passage 43 may further include at least one exhaust detection point (not shown in the accompanying drawings, please refer to the branch liquid detection point 424 for understanding), and the exhaust detection point may be connected to the exhaust valve body 431 through the exhaust channel 432. Specifically, the exhaust detection point may also be a liquid detection point provided in the exhaust passage 43. When the flow condition can be detected at the exhaust detection point: from liquid flowing to no liquid flowing, it represents that most of the sample solution has been injected into the discharge chamber 45 through the first channel 413 and the exhaust passage 43. At this time, the sample solution at the connection position between each reaction passage 42 and the first channel 413, as well as in the area in front of the connection position, has been squeezed out and emptied by the gas. In the above technical solution, by setting the exhaust detection point in the exhaust passage 43, it is possible to detect whether the sample solution at the connection position between each reaction passage 42 and the first channel 413 has been squeezed out and emptied by the gas, thereby improving the reliability of the gas isolation between each reaction passage 42, so as to improve the reliability of nucleic acid detection.

[0081] In some embodiments, the connection position between each reaction channel 42 and the first flow channel 413 is a branch connection position 4102, and the end of the first flow channel 413 for connecting with the liquid outlet 300 is a liquid inlet end 4101. The input channel 41 of the multi-channel nucleic acid reaction chip 4 may also include a pre-liquid detection point (not shown in the drawings, please refer to the branch liquid detection point 424 for understanding), which may be provided in the first flow channel 413 and located between the liquid inlet end 4101 and the branch connection position 4102 closest to the liquid inlet end 4101 (with Figure 8 For example, the branch connection position 4102 closest to the liquid inlet end 4101 is the branch connection position 4102 located at one end of the first reaction pathway 4201. In the above technical solution, by providing the pre-liquid detection point, when the sample solution flows between the piston chamber 31 and the amplification enzyme ball chamber 32 of the genetic testing kit 3, the airtightness of each pathway in the chip can be detected, thereby reducing the probability of the sample solution entering the nucleic acid amplification reaction chamber 421 without dissolving the amplification enzyme ball, and improving the reliability of the directional flow of the sample solution.

[0082] In the embodiment of the present application, the branch liquid detection point 424, the exhaust detection point and the front liquid detection point can refer to the physical groove structure on the corresponding flow channel, or can refer to a fixed position on the corresponding flow channel. The liquid detection method corresponding to the branch liquid detection point 424, the exhaust detection point and the front liquid detection point can be carried out by means of physical detection means or chemical detection means. The physical detection means can be selected from detectors such as capacitance detection, ultrasonic detection, electrode detection, infrared detection, etc., and its specific principle is: when liquid detection is performed at a fixed liquid detection point, the signal fed back by the detector when there is liquid and when there is no liquid (that is, the medium at the point changes) is different, and the difference can be used to judge whether there is liquid. The chemical detection means is to use the sample solution to react with certain chemical substances to produce obvious dominant changes, such as color changes, etc. The chemical substance placed at the liquid detection point before contacting the liquid is the original color, and after contacting the liquid, the chemical substance will turn into other colors (or, if there is liquid flowing through the liquid detection point, it will change color, and if there is no liquid flowing through the liquid detection point, it will return to its original color). Chemical detection methods can use color-changing silica gel particles, which are implanted in the liquid detection point in advance. When the reconstituted solution reaches the liquid detection point, the color-changing silica gel will change from orange to dark green (or other colors).

[0083] The embodiment of the present application integrates the valve body 44, flow channel, nucleic acid amplification reaction chamber 421, liquid detection point, and discharge chamber 45 on the same chip, making the chip function more diversified and making the nucleic acid detection results more accurate and reliable; accordingly, the genetic testing instrument 1 can automatically realize nucleic acid reaction process monitoring and closed-loop control based on the arrangement of the various valve bodies 44, nucleic acid amplification reaction chamber 421, detection points, discharge chamber 45 and other structures on the multi-channel nucleic acid reaction chip 4. For example, when the genetic testing instrument 1 receives a liquid flow signal fed back by the detector at the branch liquid detection point 424 in a certain reaction pathway 42, it will automatically control the valve control module 5 to immediately close the branch valve body 423 in the reaction pathway 42, or to close it after waiting for a period of time, that is, no longer injecting sample solution into the nucleic acid amplification reaction chamber 421 in the reaction pathway 42.

[0084] like Figure 8 As shown, in some embodiments, the connection position between each reaction channel 42 and the first flow channel 413 is a branch connection position 4102, and the end of the first flow channel 413 for connecting with the liquid outlet 300 is a liquid inlet end 4101. The multi-channel nucleic acid reaction chip 4 may further include a pre-valve body 414, which may be provided in the first flow channel 413 and located between the liquid inlet end 4101 of the first flow channel 413 and the branch connection position 4102 closest to the liquid inlet end 4101 (with a spacing of 1 / 4). Figure 8For example, the branch connection position 4102 closest to the liquid inlet end 4101 is the branch connection position 4102 located at one end of the first reaction path 4201. In the above technical solution, the provision of the pre-valve body 414 can improve the reliability of the sample solution flowing or directional flow within the designated area of ​​the multi-channel nucleic acid reaction chip 4.

[0085] After the sample to be tested completes the steps of sample lysis, nucleic acid washing, nucleic acid elution, etc. in the gene detection kit 3, the nucleic acid solution that has been purified is pumped into the piston chamber 31 of the gene detection kit 3. The PCR amplification enzyme can be made into spherical particles in advance and stored in the amplification enzyme ball chamber 32 of the kit. At this time, the nucleic acid solution can be first injected into the amplification enzyme ball chamber 32 to melt the amplification enzyme ball in the nucleic acid solution. In this step, the front valve body 414 can be closed in advance, and then the piston rod in the piston chamber 31 is pressed down. Since the front valve body 414 is in the off state, that is, the blocked state, the nucleic acid solution will enter the first branch 411 through the first liquid outlet 301, and then enter the second branch 412 through the liquid inlet end 4101 of the first flow channel 413, and then enter the amplification enzyme ball chamber 32 through the second liquid outlet 302 and melt it. After the primer ball (amplifier enzyme ball) is completely melted, the piston rod in the control piston chamber 31 is pulled upward to draw the nucleic acid solution (also referred to as the reconstitution solution or sample solution in the following embodiment) in which the PCR amplifier enzyme ball has been dissolved back into the piston chamber 31 and wait to be injected into the multi-channel nucleic acid reaction chip 4.

[0086] If the reconstitution solution needs to be injected into the multi-channel nucleic acid reaction chip 4, the piston rod in the amplification enzyme ball chamber 32 is first controlled to remain depressed, blocking the bottom channel of the amplification enzyme ball chamber 32. The pre-valve body 414 is then opened (or the branch valve body 423 in one reaction channel 42 is opened). The piston rod in the piston chamber 31 is then controlled to be depressed downward, and the reconstitution solution enters the multi-channel nucleic acid reaction chip 4 through the first liquid outlet 301. Because the bottom end of the amplification enzyme ball chamber 32 is blocked, the reconstitution solution can only flow along the first branch channel 411 and the first flow channel 413 to the pre-valve body 414. Then, through the branch connection position 4102 between the reaction channel 42 and the first flow channel 413, it flows into the branch flow channel 422 and the nucleic acid amplification reaction chamber 421.

[0087] See Figures 11 to 13 , Figure 11 This is a schematic structural diagram of the second side of the multi-channel nucleic acid reaction chip 4 shown in the second embodiment of the present application; Figure 12 This is a schematic structural diagram of a valve body 44 according to an embodiment of the present application; Figure 13 This is a schematic diagram showing the valve body 44 in the closed state according to an embodiment of the present application. Figures 6 to 13As shown, the valve body 44 in the multi-channel nucleic acid reaction chip 4 can be a branch valve body 423, an exhaust valve body 431 or a pre-valve body 414. Specifically, the valve body 44 can include an input through-hole 441, an output through-hole 442 and a stop valve cavity 440. Specifically, one end of the input through-hole 441 and one end of the output through-hole 442 are both connected to the stop valve cavity 440, and the other end of the input through-hole 441 and the other end of the output through-hole 442 are connected to the flow channel where the valve body is located (with Figure 12 For example, the other end of the input through-hole 441 of the branch valve body 423 and the other end of the output through-hole 442 are connected to the branch flow channel 422 where the branch valve body 423 is located. The stop valve chamber 440 can be squeezed by the valve core column 54 to seal one end of the input through-hole 441 and / or one end of the output through-hole 442 through the sealing membrane 402. Furthermore, the stop valve chamber 440 can be squeezed by the valve core column 54 to simultaneously seal one end of the input through-hole 441 and one end of the output through-hole 442.

[0088] In the embodiment of the present application, when the valve body 44 is in the open state, the reconstitution solution will enter the input through hole 441 through the front section flow channel (or called the upper flow channel) connected to the input through hole 441, and then be injected into the stop valve chamber 440. After the stop valve chamber 440 is filled with the reconstitution solution, the reconstitution solution enters the rear section flow channel (or called the lower flow channel) connected to the output through hole 442 through the output through hole 442, thereby realizing the circulation of the reconstitution solution in this section of the passage; when the valve body 44 is to be switched to the closed state, it is sufficient to block at least one of the input through hole 441 and the output through hole 442, that is, as long as a plug that can completely block the input through hole 441 or the output through hole 442 is used to block it (in the embodiment of the present application, the valve core column 54 is used as a plug) to block it, the current valve body can be closed.

[0089] In some embodiments, the multiple valve bodies 44 in the multi-channel nucleic acid reaction chip 4 are distributed on the same straight line. Furthermore, when the multi-channel nucleic acid reaction chip 4 is vertically inserted into one side of the genetic testing kit 3, the multiple valve bodies 44 arranged in a straight line from the bottom to the top of the multi-channel nucleic acid reaction chip 4 can be arranged in sequence as the pre-valve body 414, the branch valve body 423 in the first reaction pathway 4201, the branch valve body 423 in the second reaction pathway 4202, the branch valve body 423 in the third reaction pathway 4203, the branch valve body 423 in the fourth reaction pathway 4204, and the exhaust valve body 431. In the above technical solution, the multiple valve bodies 44 are distributed on the same straight line, which enables the corresponding valve control module 5 to achieve a simplified structure and control logic.

[0090] See Figures 14 to 17 , Figure 14 This is a schematic diagram of the overall structure of the valve control module 5 shown in one embodiment of the present application; Figure 15This is an exploded schematic diagram of a valve control module 5 according to an embodiment of the present application; Figure 16 This is a schematic diagram of a partial structure of a valve control module 5 according to an embodiment of the present application; Figure 17 FIG. 5 is a cross-sectional view of a valve control module 5 according to an embodiment of the present application. Figures 14 to 17 As shown, the valve control module 5 may include a mounting base 50, a camshaft 53, a drive motor 51, and multiple valve core columns 54. Specifically, the camshaft 53 may be rotatably connected to the mounting base 50 via a bearing; the output shaft of the drive motor 51 may be connected to the camshaft 53 via a coupling 52, and the drive motor 51 may be fixed to one end of the mounting base 50. When the multiple valve bodies 44 in the multi-channel nucleic acid reaction chip 4 are distributed along the same straight line, the mounting base 50 may also have multiple valve core column holes 501 arranged along the straight line.

[0091] Furthermore, multiple valve core posts 54 are arranged along the same straight line and housed within respective valve core post holes 501 of the mounting seat 50. The camshaft 53 includes multiple protrusions 531 (the protrusions 531 can be considered as multiple cams arranged around the outside of a cylindrical body). One end of each valve core post 54 abuts against a protrusion 531 via an elastic member 55 disposed within a valve core post hole 501. The other end of the valve core post 54 can be aligned with a valve body 44 or a non-conducting valve cavity 440 of a valve body 44. In the above technical solution, the cooperation between the protrusions 531 on the camshaft 53 and the valve core posts 54 enables the valve control module 5 to achieve on-off control of multiple valve bodies 44 using a single power source, significantly simplifying the structure and control logic of the valve control module 5.

[0092] In the embodiment of the present application, the valve control module 5 functions to direct the sample solution entering the multi-channel nucleic acid reaction chip 4 to flow in a specific sequence or flow path. Specifically, the valve control module 5 is capable of controlling the flow direction of the sample solution entering the multi-channel nucleic acid reaction chip 4. A valve core column 54 is housed within a valve core column hole 501 of the mounting base 50. Because the axes of the valve core column holes 501 are aligned, the valve core column 54 can only retract or slide out along that axis. Specifically, the valve core column holes 501 restrict the circumferential movement of the valve core column 54 while retaining its axial linear movement.

[0093] like Figure 16As shown, the camshaft 53 is provided with a plurality of protrusions 531 corresponding one to one with the valve core columns 54. One end of a valve core column 54 can be tightly attached to a protrusion 531 of the camshaft 53 via an elastic member 55. Each protrusion 531 has a boss section and a platform section. The boss section refers to the highest point on the outer surface of the protrusion 531 relative to the axis of the camshaft 53, and the platform section refers to the lowest point on the outer surface of the protrusion 531 relative to the axis of the camshaft 53. When one end of the valve core column 54 abuts against the platform section, the height of the valve core column 54 will drop a certain distance (for example, 1 mm). When one end of the valve core column 54 abuts against the boss section, the height of the valve core column 54 relative to the platform section will rise a certain distance (for example, 1 mm).

[0094] Thus, when the drive motor 51 drives the camshaft 53 to rotate, the valve stem 54 in contact with the raised portion 531 of the camshaft 53 will move upward and be pushed out when it abuts the boss section. The pushed-out valve stem 54 will squeeze the corresponding valve body 44 in the multi-channel nucleic acid reaction chip 4, closing the valve body 44 and blocking the corresponding flow path. When the camshaft 53 rotates and the valve stem 54 abuts the platform section, the valve stem 54 will retract to its original height under the action of the elastic member 55 (return spring). The valve stem 54 no longer squeezes the corresponding valve body 44 on the chip. At this time, the corresponding valve body 44 on the chip is in the open state, and the corresponding flow path is connected.

[0095] Furthermore, the camshaft 53 provided in the embodiment of the present application can reasonably design the outer contour shape of each protrusion 531 according to the control sequence of the conduction or cutoff of each valve body 44 on the chip, that is, reasonably design the relative position or relative orientation of the boss section and the platform section on each protrusion 531, so that when the camshaft 53 rotates intermittently in the specified rotation direction, each valve body 44 on the multi-channel nucleic acid reaction chip 4 can be conducted or cut off according to a preset rule.

[0096] In some embodiments, the valve control module 5 may further include an encoder, which may be located on one side of the camshaft 53. The encoder is used to monitor the rotational angle of the camshaft 53 in real time, providing feedback for the genetic testing instrument to control the drive motor 51, thereby precisely rotating the camshaft 53 to the target angle. The encoder improves the accuracy of the camshaft 53 rotation angle control, thereby precisely controlling the opening and closing of each valve body 44.

[0097] The multiple valve core columns 54 in the valve control module 5 can correspond one-to-one with the multiple valve bodies 44 on the multi-channel nucleic acid reaction chip 4. For example, six valve core columns 54 correspond to one pre-valve body 414, four branch valve bodies 423, and one exhaust valve body 431, respectively. The on / off control logic of the six valve bodies 44 is shown in Table 1 below, where the first branch valve body refers to the branch valve body 423 in the first reaction pathway 4201, the second branch valve body refers to the branch valve body 423 in the second reaction pathway 4202, the third branch valve body refers to the branch valve body 423 in the third reaction pathway 4203, and the fourth branch valve body refers to the branch valve body 423 in the fourth reaction pathway 4204.

[0098] In addition, in the case where the pre-valve body 414 is not provided, in the step of redissolving the amplification enzyme ball, all valve bodies 44 in the multi-channel nucleic acid reaction chip 4 can also be closed first. When the piston rod in the control piston chamber 31 is pressed down, since each reaction channel 42 is in a blocked state, the nucleic acid solution will enter the amplification enzyme ball chamber 32 along the first branch 411 and the second branch 412 and melt it. Although a small amount of nucleic acid solution may enter the first flow channel 413, the small amount of residual liquid in the chip can be sucked back into the piston chamber 31 by pulling the piston rod in the piston chamber 31. Then, when the redissolving solution is injected into each nucleic acid amplification reaction chamber 421, the branch valve bodies 423 in the first reaction channel 4201 to the fourth reaction channel 4204 can be controlled to be connected and closed in sequence to improve the uniformity of the liquid in each reaction chamber.

[0099] Table 1 - Control logic of each valve body 44 in the multi-channel nucleic acid reaction chip 4:

[0100]

[0101] In the embodiment of the present application, a multi-channel nucleic acid reaction chip 4 may include four nucleic acid amplification reaction chambers 421 (reaction pathways 42). The number of reaction pathways 42 and nucleic acid amplification reaction chambers 421 can be set according to actual detection requirements, and can be 2, 4, 8, 16, 32, etc.

[0102] Please combine Figures 6 to 10As shown, each nucleic acid amplification reaction chamber 421 includes a reaction inlet 4211 and a reaction outlet 4212. Furthermore, when the multi-channel nucleic acid reaction chip 4 is plugged into the gene detection kit 3 in an upright state, the reaction inlet 4211 can be located at the bottom end of the nucleic acid amplification reaction chamber 421, and the reaction outlet 4212 can be located at the top end of the nucleic acid amplification reaction chamber 421. In the above technical solution, the reaction inlet 4211 is located at the bottom end of the nucleic acid amplification reaction chamber 421, and the reaction outlet 4212 is located at the top end of the nucleic acid amplification reaction chamber 421, so that the nucleic acid amplification reaction chamber 421 is more easily filled with the sample solution, reducing the probability of bubbles remaining in the nucleic acid amplification reaction chamber 421, thereby improving the uniformity of the sample solution in each nucleic acid amplification reaction chamber 421.

[0103] In some embodiments, the centers of the multiple nucleic acid amplification reaction chambers 421 can be located on the same straight line. The above technical solution makes the structural layout of the multi-channel nucleic acid reaction chip 4 more reasonable and can also make the overall structure of the corresponding fluorescence collection and analysis module 6 more compact and reasonable.

[0104] In some embodiments, the multi-channel nucleic acid reaction chip 4 may further include at least one partitioning space 46, which is typically provided between two adjacent nucleic acid amplification reaction chambers 421. In the above technical solution, by providing the partitioning space 46, the multi-channel nucleic acid reaction chip 4 can reduce fluorescence crosstalk between adjacent nucleic acid amplification reaction chambers 421 by means of light refraction and scattering, thereby improving the accuracy and reliability of nucleic acid detection.

[0105] See Figures 18 and 19 , Figure 18 This is a schematic structural diagram of a fluorescence collection and analysis module 6 according to an embodiment of the present application; Figure 19 FIG. 4 is a schematic diagram showing a multi-channel nucleic acid reaction chip 4 loaded with a fluorescence collection and analysis module 6 according to an embodiment of the present application. Figures 18 and 19 As shown, the fluorescence collection and analysis module 6 includes a fixing frame 61 having a fluorescence collection hole 60 . The fluorescence collection hole 60 is used to accommodate one end of the multi-channel nucleic acid reaction chip 4 having a nucleic acid amplification reaction chamber 421 .

[0106] Fluorescence acquisition and analysis module 6 can utilize commonly used or general fluorescence detection equipment in the field of molecular diagnostics. Its specific principle is: nucleic acids are amplified under the constraints of specific primers. Because the primers are designed with specific binding sites, the free gene produces a specific fluorescence when it binds specifically to the site. As the PCR reaction continues, the fluorescent group gradually increases in intensity. Therefore, the fluorescence signal is collected and analyzed during the PCR reaction. Fluorescence acquisition and analysis module 6 collects and analyzes the fluorescence signal simultaneously, and outputs a report based on the detected biological sample through data analysis.

[0107] The multi-channel nucleic acid reaction chip 4 provided in the embodiment of the present application enables multiple nucleic acid amplification reaction chambers 421 to simultaneously perform polymerase chain reactions, thereby improving detection efficiency. In addition, through the coordination of the valve control module 5 and the liquid detection points, the genetic testing instrument 1 can accurately and efficiently control the flow path and liquid flow direction in the chip, enabling the entire nucleic acid detection and analysis system (genetic testing instrument 1) to operate efficiently and stably, and output more accurate detection results.

[0108] Please combine Figures 1 to 19 As shown, the application method of the gene detection device 2 is as follows:

[0109] First, the operator obtains a genetic testing kit 3 capable of performing nucleic acid extraction and purification. A multi-channel nucleic acid reaction chip 4 is connected to one side of the kit. The operator removes the screw cap 30 on top of the kit, adds the biological sample to be tested, and replaces and tightens the screw cap 30. The operator then places the sample-added genetic testing kit 3 into the genetic testing instrument 1. Subsequent steps, such as sample lysis, nucleic acid washing, and nucleic acid elution, are automatically controlled by the genetic testing instrument 1.

[0110] After the nucleic acid sample completes the steps of sample lysis, nucleic acid washing, and nucleic acid elution in the gene detection kit 3, it will be drawn into the piston chamber 31 of the gene detection kit 3 and the step of re-dissolving the PCR amplification enzyme ball will begin. The gene detection instrument 1 can first control the valve control module 5 to close the pre-valve body 414 in the multi-channel nucleic acid reaction chip 4, or the gene detection instrument 1 can first control the valve control module 5 to close all valve bodies 44 in the multi-channel nucleic acid reaction chip 4, and then control the piston rod in the piston chamber 31 in the gene detection kit 3 to press down. Since the flow channel below the piston chamber 31 leading to each reaction channel is in a blocked state, the nucleic acid solution will enter the amplification enzyme ball chamber 32 along the first branch 411 and the second branch 412 and melt it. After the PCR amplification enzyme ball is completely melted, the piston rod in the piston chamber 31 is pulled upward, and the re-solution that has dissolved the PCR amplification enzyme ball can be drawn back into the piston chamber 31, waiting to be injected into the multi-channel nucleic acid reaction chip 4.

[0111] Then, the reconstitution solution is injected into the multi-channel nucleic acid reaction chip 4. First, the piston rod in the amplification enzyme ball chamber 32 should be controlled to be continuously pressed down to block the flow channel at the bottom of the amplification enzyme ball chamber 32, and then the front valve body 414 should be opened, or the branch valve body 423 in a reaction path 42 should be opened. At this time, the piston rod in the piston chamber 31 is pressed down again, and the reconstitution solution will enter the multi-channel nucleic acid reaction chip 4 and flow into the nucleic acid amplification reaction chamber 421 through the first flow channel 413.

[0112] During the flow of the reconstitution solution into the nucleic acid amplification reaction chamber 421 via the first flow channel 413, it may split into two paths. One path of the reconstitution solution may flow slowly or not at all along the vertical first flow channel 413 (because the exhaust valve body 431 is closed), while the other path of the reconstitution solution will flow along the branch flow channel 422 where an open branch valve body 423 is located, entering the nucleic acid amplification reaction chamber 421 in the reaction channel 42. When the reconstitution solution reaches the branch liquid detection point 424, the valve control module 5 controls the branch valve body 423 to close immediately or after a specified period of time, thereby blocking the reaction channel 42 and preventing further liquid injection. From the bottom to the top of the multi-channel nucleic acid reaction chip 4, the valve control module 5 can sequentially open and close the branch valve bodies 423 in each reaction channel 42, in the order of the first reaction channel 4201, the second reaction channel 4202, the third reaction channel 4203, and the fourth reaction channel 4204, from bottom to top. After the nucleic acid amplification reaction chamber 421 in the current reaction channel is filled with the reconstitution solution, the branch valve body 423 in the current reaction channel is closed and the branch valve body 423 in the next reaction channel 42 is opened. In this way, each nucleic acid amplification reaction chamber 421 on the chip is filled in turn.

[0113] After all nucleic acid amplification reaction chambers 421 are filled with sample solution, the genetic testing instrument 1 opens the exhaust valve body 431 and continues to control the piston rod in the piston chamber 31 to press down, so that the excess solution and bubbles in the multi-channel nucleic acid reaction chip 4 are all injected into the exhaust chamber through the first flow channel 413 and the exhaust passage 43, so as to maintain the cleanliness and gas sealing state in the first flow channel 413, effectively reduce liquid crosstalk between the nucleic acid amplification reaction chambers 421, and improve the accuracy of the test results.

[0114] Finally, during the PCR amplification reaction based on the multi-channel nucleic acid reaction chip 4, the fluorescence collection and analysis module 6 will collect fluorescence signals at the fluorescence signal collection points in each nucleic acid amplification reaction chamber 421, and then analyze the collected fluorescence signals and output a report.

[0115] The above description is merely a preferred embodiment of the present application and is not intended to limit the present application. Various modifications and variations are possible for those skilled in the art. Any modifications, equivalent substitutions, or improvements made within the spirit and principles of the present application shall be included within the scope of protection of the present application.

Claims

1. A multi-channel nucleic acid reaction chip for connecting to a gene detection kit, characterized in that: The multi-channel nucleic acid reaction chip comprises: a first flow channel, one end of which is connected to the liquid outlet of the gene detection kit; a plurality of independent reaction pathways, one end of each reaction pathway being in communication with the first flow channel, the reaction pathways comprising a nucleic acid amplification reaction chamber and a branch valve body, the nucleic acid amplification reaction chamber being in communication with the branch valve body; An exhaust passage includes an exhaust valve body, and the exhaust valve body is communicated with the other end of the first flow channel.

2. The multi-channel nucleic acid reaction chip according to claim 1, characterized in that: The reaction pathway further includes at least one branch liquid detection point, which is connected to the nucleic acid amplification reaction chamber through a branch flow channel, or the branch liquid detection point is arranged in the nucleic acid amplification reaction chamber.

3. The multi-channel nucleic acid reaction chip according to claim 1, characterized in that: The exhaust passage further includes at least one exhaust detection point, and the exhaust detection point is connected to the exhaust valve body through an exhaust flow channel.

4. The multi-channel nucleic acid reaction chip according to claim 1, characterized in that: The connection position between each of the reaction channels and the first flow channel is a branch connection position, and the end of the first flow channel used for connecting with the liquid outlet is a liquid inlet end; The multi-channel nucleic acid reaction chip further includes a front liquid detection point, which is provided in the first flow channel and is located between the liquid inlet end and the branch connection position closest to the liquid inlet end.

5. The multi-channel nucleic acid reaction chip according to claim 1, characterized in that: The connection position between each of the reaction channels and the first flow channel is a branch connection position, and the end of the first flow channel used for connecting with the liquid outlet is a liquid inlet end; The multi-channel nucleic acid reaction chip further includes a pre-valve body, which is provided in the first flow channel and located between the liquid inlet end and the branch communication position closest to the liquid inlet end.

6. The multi-channel nucleic acid reaction chip according to any one of claims 1 to 5, characterized in that: The gene detection kit includes a first liquid outlet and a second liquid outlet, and the multi-channel nucleic acid reaction chip also includes a first branch and a second branch; The first branch is connected to the second branch at the liquid inlet end of the first flow channel. The first branch is used to communicate with the first liquid outlet, and the second branch is used to communicate with the second liquid outlet.

7. The multi-channel nucleic acid reaction chip according to any one of claims 1 to 5, characterized in that: The valve body in the multi-channel nucleic acid reaction chip includes: An input through hole, an output through hole and a stop valve cavity, one end of the input through hole and one end of the output through hole are both connected to the stop valve cavity, and the other end of the input through hole and the other end of the output through hole are connected to the flow channel; the stop valve cavity can block one end of the input through hole and / or one end of the output through hole by squeezing the valve core column.

8. The multi-channel nucleic acid reaction chip according to claim 7, characterized in that: The multiple valve bodies in the multi-channel nucleic acid reaction chip are distributed on the same straight line.

9. The multi-channel nucleic acid reaction chip according to any one of claims 1 to 5, characterized in that: The multi-channel nucleic acid reaction chip further includes a plurality of exhaust cavities, which are connected to the reaction passages and / or the exhaust passages.

10. The multi-channel nucleic acid reaction chip according to claim 1, characterized in that: The multi-channel nucleic acid reaction chip further comprises at least one separation space, which is provided between two adjacent nucleic acid amplification reaction chambers.

11. The multi-channel nucleic acid reaction chip according to any one of claims 1 to 5 and 10, characterized in that: The center points of the multiple nucleic acid amplification reaction chambers are located on the same straight line.

12. The multi-channel nucleic acid reaction chip according to any one of claims 1 to 5 and 10, characterized in that: The nucleic acid amplification reaction chamber includes a reaction inlet and a reaction outlet; When the multi-channel nucleic acid reaction chip is plugged into the gene detection kit in an upright state, the reaction inlet is located at the bottom end of the nucleic acid amplification reaction chamber, and the reaction outlet is located at the top end of the nucleic acid amplification reaction chamber.

13. The multi-channel nucleic acid reaction chip according to any one of claims 1 to 5 and 10, characterized in that: The multi-channel nucleic acid reaction chip comprises a substrate and a sealing film, wherein the substrate has a first surface and a second surface disposed opposite to each other; Grooves and / or through holes are formed on the first surface and / or the second surface, and the sealing film is respectively covered on the first surface and the second surface to cooperate with the substrate to form the first flow channel, the nucleic acid amplification reaction chamber, the branch valve body and the exhaust valve body.

14. The multi-channel nucleic acid reaction chip according to any one of claims 1 to 5 and 10, characterized in that: The multi-channel nucleic acid reaction chip includes a chip body and a plug-in interface, the first flow channel is provided in the chip body; the plug-in interface is communicated with the first flow channel and connected to the chip body, and the plug-in interface is used to dock with the liquid outlet.

15. A gene detection device, characterized in that: The gene detection device comprises: A gene detection kit, wherein the gene detection kit has a liquid outlet; The multi-channel nucleic acid reaction chip according to any one of claims 1 to 14, wherein one end of the multi-channel nucleic acid reaction chip is connected to the gene detection kit at the liquid outlet; a valve control module, the valve control module being arranged on one side of the multi-channel nucleic acid reaction chip and being arranged adjacent to each valve body of the multi-channel nucleic acid reaction chip; The fluorescence collection and analysis module has a fluorescence collection hole, and the multi-channel nucleic acid reaction chip has one end of the nucleic acid amplification reaction chamber, which extends into and is accommodated in the fluorescence collection hole.

16. The gene detection device according to claim 15, characterized in that When the plurality of valve bodies in the multi-channel nucleic acid reaction chip are distributed on the same straight line, the valve control module includes: a mounting seat and a camshaft, wherein the camshaft is rotatably connected to the mounting seat; a driving motor, wherein an output shaft of the driving motor is connected to the camshaft; Multiple valve core columns are arranged along the same straight line and accommodated in each valve core column hole of the mounting seat, and the camshaft includes multiple protrusions; one end of the valve core column rests on the protrusion, and the other end of the valve core column is aligned with the valve body.

17. A gene detection instrument, characterized in that: The gene detection instrument includes the gene detection device according to claim 15 or 16.

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