Nucleic acid detection card box and nucleic acid detection method

Abstract

The invention relates to the technical field of in-vitro medical diagnosis, and discloses a nucleic acid detection card box and a nucleic acid detection method.The card box is provided with a sample inlet, and a sample cavity communicated with the sample inlet is formed in the card box; a cracking cavity, an elution cavity and a reaction cavity which are sequentially communicated through a flow channel with a valve are further formed in the clamping box, and the sample cavity is communicated with the cracking cavity through the flow channel; a magnetic bead cavity is further formed in the clamping box, and the magnetic bead cavity communicates with the cracking cavity through a flow channel; a waste liquid cavity is also formed in the clamping box, and is respectively communicated with the cracking cavity and the elution cavity through a flow channel with a valve; a first air hole communicated with at least one of the sample cavity and the cracking cavity, a second air hole communicated with the elution cavity and a reagent inlet respectively communicated with the cracking cavity and the elution cavity are formed in the clamping box. Through the mode, the detection efficiency and the accuracy of a detection result can be improved, and meanwhile it is guaranteed that the card box is small in size, simple in structure and low in cost.

Description

TECHNICAL FIELD

[0001] The embodiment of the present application relates to the technical field of in vitro medical diagnosis, in particular to a nucleic acid detection card box and a nucleic acid detection method. BACKGROUND

[0002] Nucleic acid detection is widely used in the field of biological medicine and has high clinical significance, especially in the field of in vitro medical diagnosis. Compared with traditional tube-type biological analysis, the detection card box has the characteristics of small reaction volume and can save reagent consumption.

[0003] The nucleic acid detection technology generally includes the steps of lysis, combination, washing, elution, and subsequent PCR (polymerase chain reaction, also known as in vitro DNA amplification technology) and optical detection.

[0004] In the existing nucleic acid detection card box, some have low integration, many processes need manual intervention, so the detection efficiency is low, the probability of misoperation is high, the detection result is unstable, and the accuracy cannot be guaranteed, and some have high integration, but the number of parts is large, the structure is complex, and the cost is high. SUMMARY

[0005] In view of the above problems, the embodiment of the present application provides a nucleic acid detection card box and a nucleic acid detection method, which can improve the detection efficiency and the accuracy of the detection result while ensuring that the card box has a small volume, a simple structure and a low cost.

[0006] According to an aspect of the embodiment of the present application, a nucleic acid detection card box is provided, the card box is provided with a sample inlet, and the inside of the card box is provided with a sample chamber in communication with the sample inlet; the inside of the card box is further provided with a lysis chamber, an elution chamber and a reaction chamber which are sequentially communicated through flow channels with valves, and the sample chamber is communicated with the lysis chamber through a flow channel; the inside of the card box is further provided with a magnetic bead chamber, and the magnetic bead chamber contains magnetic beads, and the magnetic bead chamber is communicated with the lysis chamber through a flow channel; the inside of the card box is further provided with a waste liquid chamber, and the waste liquid chamber is communicated with the lysis chamber and the elution chamber through flow channels with valves; the card box is provided with a first air hole communicated with at least one of the sample chamber and the lysis chamber; the card box is further provided with a second air hole communicated with the elution chamber; and the card box is further provided with a reagent inlet communicated with the lysis chamber and the elution chamber.

[0007] In an optional manner, the magnetic bead chamber is communicated between the sample chamber and the lysis chamber through a flow channel, so that the sample chamber is communicated with the lysis chamber through a flow channel and the magnetic bead chamber.

[0008] In an optional manner, the first air hole is communicated with the sample chamber, and the first air hole is used to drive the sample from the sample chamber into the magnetic bead chamber when a positive pressure is introduced.

[0009] In an alternative way, the reaction chamber is further communicated with the waste liquid chamber through a flow channel with a water-proof and air-permeable membrane, so that in the process of the liquid in the elution chamber entering the reaction chamber, the gas in the reaction chamber is discharged to the waste liquid chamber, the liquid is blocked inside the reaction chamber, and the aerosol in the reaction chamber is prevented from leaking.

[0010] In an alternative way, a blocking valve is arranged in the flow channel between the reaction chamber and the elution chamber and in the flow channel between the reaction chamber and the waste liquid chamber, and the blocking valve is configured to be closed after the liquid injection in the reaction chamber is completed.

[0011] In an alternative way, the reaction reagent is contained in the interior of the reaction chamber, and the reaction reagent is used to react with the liquid injected from the elution chamber into the reaction chamber.

[0012] In an alternative way, one side of the cartridge is formed with a sheet structure, the reaction chamber is arranged in the interior of the sheet structure, and the wall thickness of the sheet structure is smaller than that of the rest of the cartridge.

[0013] In an alternative way, the reaction chamber is multiple, and the multiple reaction chambers are arranged in parallel.

[0014] In an alternative way, the reagent inlet includes a first inlet and a second inlet which are respectively communicated with the lysis chamber, the first inlet is used for the lysis reagent and the binding reagent to enter the lysis chamber, and the second inlet is used for the washing reagent to enter the lysis chamber; the reagent inlet further includes a third inlet and a fourth inlet, the third inlet is used for the washing reagent to enter the elution chamber, and the fourth inlet is used for the elution reagent to enter the elution chamber.

[0015] In an alternative way, the cartridge is further provided with a pressure relief hole which is communicated with the waste liquid chamber.

[0016] In an alternative way, at least one of the sample chamber and the lysis chamber is covered with a water-proof and air-permeable membrane between the first air hole, the elution chamber between the second air hole, and the pressure relief hole between the waste liquid chamber, and the water-proof and air-permeable membrane is used to prevent the aerosol and the vaporized reagent inside from leaking.

[0017] According to another aspect of the embodiments of the present application, a nucleic acid detection method is provided, which is applied to the nucleic acid detection cartridge in any of the above embodiments. The method comprises: injecting a sample into the sample chamber through the sample inlet; keeping all valves closed; introducing pressure into the first air hole to make the sample flow into the magnetic bead chamber to mix with the magnetic beads to form a first mixed fluid; introducing pressure into the first air hole again to make the first mixed fluid flow into the lysis chamber; injecting a lysis agent and a binding agent into the lysis chamber through the reagent inlet connected with the lysis chamber, so that the lysis agent and the binding agent mix and react with the first mixed fluid to form a second mixed fluid, wherein the cells and / or viruses in the first mixed fluid are lysed under the action of the lysis agent to expose the nucleic acid, and the nucleic acid is combined with the magnetic beads under the action of the binding agent; adsorbing the magnetic beads in the second mixed fluid to fix the magnetic beads combined with the nucleic acid in the lysis chamber, opening the first valve on the flow channel between the lysis chamber and the waste liquid chamber, and introducing positive pressure into the first air hole to make the liquid in the lysis chamber flow into the waste liquid chamber, then closing the first valve and releasing the adsorption of the magnetic beads; injecting a washing agent into the lysis chamber through the reagent inlet connected with the lysis chamber to clean the magnetic beads and form a third mixed fluid; opening the second valve on the flow channel between the lysis chamber and the elution chamber, and introducing positive pressure into the first air hole or negative pressure into the second air hole to make the third mixed fluid flow into the elution chamber, then closing the second valve; adsorbing the magnetic beads in the third mixed fluid to fix the magnetic beads in the elution chamber, opening the third valve on the flow channel between the elution chamber and the waste liquid chamber, and introducing positive pressure into the first air hole or the second air hole to make the liquid in the elution chamber flow into the waste liquid chamber, then closing the third valve and releasing the adsorption of the magnetic beads; injecting a washing agent into the elution chamber through the reagent inlet connected with the elution chamber to clean the magnetic beads and form a fourth mixed fluid; adsorbing the magnetic beads in the fourth mixed fluid to fix the magnetic beads in the elution chamber, opening the third valve, and introducing positive pressure into the first air hole or the second air hole to make the liquid in the elution chamber flow into the waste liquid chamber, then closing the third valve and releasing the adsorption of the magnetic beads; opening the second valve and / or the third valve, and drying the magnetic beads in the elution chamber to make the liquid on the surface of the magnetic beads and the liquid in the elution chamber vaporize and then condense in the lysis chamber and / or the waste liquid chamber, then closing the second valve and / or the third valve; injecting an elution agent into the elution chamber through the reagent inlet connected with the elution chamber to elute the nucleic acid from the magnetic beads; adsorbing the magnetic beads in the elution chamber to fix the magnetic beads in the elution chamber, opening the fourth valve on the flow channel between the elution chamber and the reaction chamber, and introducing positive pressure into the first air hole or the second air hole to inject the liquid mixed with the nucleic acid in the elution chamber into the reaction chamber for detection.

[0018] In an alternative mode, after the step of injecting the lysis reagent and the binding reagent into the lysis chamber through the reagent inlet in communication with the lysis chamber, mixing and reacting the lysis reagent and the binding reagent with the first mixed fluid to form the second mixed fluid, and / or, after the step of injecting the washing reagent into the lysis chamber through the reagent inlet in communication with the lysis chamber to wash the magnetic beads and form the third mixed fluid, and / or, after the step of opening the second valve on the flow channel between the lysis chamber and the elution chamber, and introducing positive pressure through the first air hole or negative pressure through the second air hole to make the third mixed fluid flow to the elution chamber, and then closing the second valve, and / or, after the step of injecting the washing reagent into the elution chamber through the reagent inlet in communication with the elution chamber to wash the magnetic beads and form the fourth mixed fluid, and / or, after the step of injecting the elution reagent into the elution chamber through the reagent inlet in communication with the elution chamber to elute the nucleic acid from the magnetic beads, the method further comprises the step of mixing the fluid.

[0019] The cartridge provided by the embodiments of the present application is based on the research result that lysis and binding can be performed synchronously and do not affect each other. Therefore, only the lysis chamber is arranged in the cartridge to be responsible for lysis of the nucleic acid and binding with the magnetic beads. In order to prevent the impurities possibly remaining on the inner wall of the lysis chamber from affecting the detection result, the elution chamber is further arranged to be responsible for the step of eluting the nucleic acid from the magnetic beads, so as to ensure the extraction purity of the nucleic acid in the liquid after elution, improve the accuracy of the detection result, and reduce the volume and cost of the cartridge as much as possible. The waste liquid chambers in communication with the lysis chamber and the elution chamber respectively are further arranged to ensure the orderly discharge of the waste liquid in the lysis chamber and the elution chamber. Further, the valves on the flow channels between the chambers and the air holes in communication with part of the chambers are arranged, so that the detection process is simple to operate and the internal fluid flow direction can be accurately controlled. At the same time, the temperature difference between the elution chamber and the lysis chamber and / or the waste liquid chamber and the control of the valves therebetween can be utilized, so that when the magnetic beads are dried, the vaporized gas can flow into the lysis chamber and / or the waste liquid chamber to be condensed, so as not to flow back into the elution chamber, and the purity of the nucleic acid extraction is further improved.

[0020] A key factor to ensure the accuracy of nucleic acid detection is to avoid the mixture of impurities generated by lysis into the solution after nucleic acid elution, because the solution after nucleic acid elution needs to be injected into the reaction chamber for PCR amplification detection, if the eluted solution mixes with the impurities generated by lysis, it will affect the accuracy of PCR amplification detection. It is for this reason that in the cartridge provided in the embodiments of the present application, the lysis chamber and the elution chamber are first independently arranged, the lysis chamber is responsible for lysis, and the elution chamber is responsible for elution. Compared with the same cavity that performs lysis and elution, it can prevent the impurities generated by lysis from affecting the elution, and through multiple washing, it can well avoid the mixture of impurities generated by lysis into the solution after elution.

[0021] In addition, the cartridge provided in the embodiments of the present application also sets up independent air holes for lysis and elution, namely first air hole and second air hole, the first air hole is in communication with at least one of the sample chamber and the lysis chamber, mainly responsible for driving the solution flow between the sample chamber and the lysis chamber, and the second air hole is in communication with the elution chamber, responsible for driving the solution flow in the elution chamber to the waste chamber and the reaction chamber. Compared with the driving mode of setting only one air hole and driving the solution flow by the same air hole, it can prevent the impurities remaining in the lysis chamber from mixing into the elution chamber during liquid driving, so as to avoid the pollution of the extraction product (i.e. the detection solution containing nucleic acid obtained after the magnetic bead elution) in the elution chamber, and prevent the subsequent nucleic acid detection result from being disturbed.

[0022] The above description is only a summary of the technical solutions of the present application, in order to more clearly understand the technical means of the present application, which can be implemented according to the content of the specification, and in order to make the above and other purposes, characteristics and advantages of the present application more obvious and easy to understand, the following specific embodiments of the present application are described. BRIEF DESCRIPTION OF DRAWINGS

[0023] Various other advantages and benefits will become apparent to those of ordinary skill in the art upon reading the following detailed description of the preferred embodiments. The accompanying drawings are intended to depict only preferred embodiments of the application, and therefore should not be considered to limit the scope of the application in any way. Similarly, like reference numerals are intended to represent like parts throughout the various figures. In the drawings:

[0024] Figure 1 The perspective view of the nucleic acid detection cartridge provided in the embodiments of the present application is shown from one angle;

[0025] Figure 2 The perspective view of the nucleic acid detection cartridge provided in the embodiments of the present application is shown from another angle;

[0026] Figure 3 The exploded view of the nucleic acid detection cartridge provided in the embodiments of the present application is shown from one angle;

[0027] Figure 4 An exploded structural schematic diagram of the nucleic acid detection card provided by the embodiment of the present application from another perspective;

[0028] Figure 5 An internal perspective structural schematic diagram of the nucleic acid detection card provided by the embodiment of the present application from a front perspective;

[0029] Figure 6 An exploded structural schematic diagram of the nucleic acid detection card provided by another embodiment of the present application from a perspective; Figure 4 An enlarged structural schematic diagram at A;

[0030] Figure 7 An exploded structural schematic diagram of the nucleic acid detection card provided by another embodiment of the present application from a perspective;

[0031] Figure 8 An exploded structural schematic diagram of the nucleic acid detection card provided by another embodiment of the present application from another perspective;

[0032] Figure 9 An internal perspective structural schematic diagram of the nucleic acid detection card provided by another embodiment of the present application from a side plane perspective;

[0033] Figure 10 A structural schematic diagram of the nucleic acid detection card provided by another embodiment of the present application from another side plane perspective;

[0034] Figure 11 A structural schematic diagram of the nucleic acid detection card provided by the embodiment of the present application from a bottom perspective;

[0035] Figure 12 A flow schematic diagram of the nucleic acid detection method provided by the embodiment of the present application;

[0036] Figure 13 A flow schematic diagram of the nucleic acid detection method provided by another embodiment of the present application.

[0037] The reference signs in the detailed description of the embodiments are as follows:

[0038] 100, card box; 101, card box body; 102, cover piece;

[0039] 110, sample inlet; 111, sealing cover; 120, sample cavity; 130, magnetic bead cavity; 140, lysis cavity; 150, elution cavity; 160, reaction cavity; 161, waterproof and breathable membrane; 162, second through hole; 163, blocking valve; 170, waste liquid cavity; 171, water absorption piece; 180, sheet structure;

[0040] 11, first flow channel; 12, second flow channel; 121, serpentine flow channel; 13, third flow channel; 14, fourth flow channel; 15, fifth flow channel; 16, sixth flow channel; 17, seventh flow channel;

[0041] 21, first valve; 22, second valve; 23, third valve; 24, fourth valve; 25, first through hole; 26, stepped surface; 27, pressing piece;

[0042] 31, first air hole; 32, second air hole;

[0043] 41, first inlet; 42, second inlet; 43, third inlet; 44, fourth inlet;

[0044] 51, first reagent package; 52, second reagent package; 53, third reagent package; 54, fourth reagent package;

[0045] 60, pressure relief hole;

[0046] 71, first waterproof and air permeable membrane; 72, second waterproof and air permeable membrane; 73, third waterproof and air permeable membrane;

[0047] 81, first water-absorbing and air permeable membrane; 82, second water-absorbing and air permeable membrane;

[0048] 91, first channel; 92, second channel; 921, micro valve. DETAILED DESCRIPTION

[0049] The embodiments of the technical scheme of the present application will be described in detail below with reference to the drawings. The following embodiments are only used to more clearly illustrate the technical scheme of the present application, and therefore only serve as examples, and cannot limit the protection scope of the present application.

[0050] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs; the terms used herein are only for the purpose of describing specific embodiments and are not intended to limit the present application; the terms "include" and "have" and any variations thereof in the specification and claims of the present application and the above description of drawings are intended to cover non-exclusive inclusion.

[0051] In the description of the embodiments of the present application, the technical terms "first", "second", etc. are only used to distinguish different objects, and cannot be understood as indicating or implying relative importance or implicitly indicating the number, specific order or primary and secondary relationship of the indicated technical features. In the description of the embodiments of the present application, the meaning of "a plurality of" is two or more, unless otherwise explicitly specified.

[0052] Reference herein to an "embodiment" means that a particular feature, structure, or characteristic described in connection with the embodiment can be included in at least one embodiment of the application. The appearances of the phrase in various places in the specification are not necessarily all referring to the same embodiment, nor are they necessarily mutually exclusive of one another. It is expressly understood that the embodiments described herein are merely examples from among a great variety of embodiments that can be made in light of the present disclosure.

[0053] In the description of the embodiments of the present application, the term "and / or" is merely an association relationship of the associated objects, which means that there can be three relationships, for example, A and / or B, which can mean that A exists, A and B exist, and B exists. In addition, the character " / " herein generally represents that the front and rear associated objects are in an "or" relationship.

[0054] In the description of the embodiments of the present application, the term "a plurality of" refers to two or more (including two), and similarly, "a plurality of groups" refers to two or more groups (including two groups), and "a plurality of pieces" refers to two or more pieces (including two pieces).

[0055] In the description of the embodiments of the present application, the technical terms "center", "longitudinal", "transverse", "length", "width", "thickness", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", "clockwise", "counterclockwise", "axial", "radial", "circumferential", and the like indicate the orientation or positional relationship shown in the drawings, and are only for the convenience of describing the embodiments of the present application and simplifying the description, and are not intended to indicate or imply that the devices or elements referred to must have a particular orientation, be constructed and operated in a particular orientation, and therefore cannot be understood as a limitation on the embodiments of the present application.

[0056] In the description of the embodiments of the present application, unless otherwise explicitly specified and limited, the technical terms "mounting", "connection", "connection", "fixing", and the like should be understood in a broad sense, for example, can be fixedly connected, or can be detachably connected, or can be integrated; can be mechanically connected, or can be electrically connected; can be directly connected, or can be indirectly connected through an intermediate medium; can be the internal communication of two elements or the interaction relationship between two elements. For those skilled in the art, the specific meaning of the above terms in the embodiments of the present application can be understood according to the specific circumstances.

[0057] Compared with traditional tube type biological analysis, nucleic acid detection cartridges, such as microfluidic cartridges, not only have the characteristics of small reaction volume, saving reagent consumption, etc., but also have the characteristics of small pollution, easy to realize high-throughput analysis, low cost, strong adaptability, small size, flexible design space, fast detection speed, etc., and can disperse a complex system into a large number of simple systems containing only a single research object, so as to make the background simple and facilitate quantitative and qualitative analysis.

[0058] For some high-integration and full-featured cartridges, in the design, the internal generally need to set up the corresponding chamber and flow channel for different operation steps, respectively, which will lead to the number of integrated chambers and flow channels in the cartridge is very large, resulting in the cartridge volume is large, the structure is complex.

[0059] To ensure the miniaturization of the cartridge product, different reactions in the nucleic acid detection process can be considered in the same chamber to reduce the number of chambers, and thus achieve the purpose of reducing the volume of the cartridge. However, when different treatments are performed on the sample in the same chamber, the reactions may affect each other, resulting in a large number of impurities in the final detection liquid, so as to ensure the sample processing capacity and the accuracy of the detection result.

[0060] In view of the above problems, the inventors of the present application analyze the processing characteristics of each step in the nucleic acid detection process, and then design the internal structure of the cartridge. By designing the communication relationship between the chamber, the reagent inlet, the air hole and the flow channel, and the position of the valve that can control the opening and closing of the flow channel, the whole process of lysis, combination, washing, elution and PCR detection can be ensured, and the processing steps will not affect each other, and the detection efficiency and the accuracy of the detection result are high. At the same time, the number of chambers and flow channels required is minimized, thereby ensuring that the cartridge has a smaller volume and lower processing cost.

[0061] According to an aspect of an embodiment of the present application, a nucleic acid detection cartridge is provided, which will be described in detail with reference to Figures 1 to 4 , wherein Figure 1 and Figure 2 show the perspective structure of the nucleic acid detection cartridge from two angles, respectively, Figure 3 and Figure 4 show the exploded structure of the nucleic acid detection cartridge from two angles, respectively. As shown in the figure, the cartridge 100 is provided with a sample inlet 110, and the internal of the cartridge 100 is provided with a sample chamber 120 and a magnetic bead chamber 130 which are communicated by flow channels, in particular, as shown in the perspective view of Figure 4 and the planar view of Figure 5 , the sample chamber 120 and the magnetic bead chamber 130 are communicated by a first flow channel 11, the sample chamber 120 is also communicated with the sample inlet 110, and the magnetic bead chamber 130 contains magnetic beads (not shown in the figure).

[0062] Please refer to Figures 3 to 5The cartridge 100 also contains a pyrolysis chamber 140, an elution chamber 150, and a reaction chamber 160, which are sequentially connected by flow channels with valves. The pyrolysis chamber 140 is also connected to the magnetic bead chamber 130 through a flow channel. Specifically, the magnetic bead chamber 130 is connected to the pyrolysis chamber 140 through a second flow channel 12, the pyrolysis chamber 140 is connected to the elution chamber 150 through a third flow channel 13 with a second valve 22, and the elution chamber 150 is connected to the reaction chamber 160 through a fourth flow channel 14 with a fourth valve 24.

[0063] Furthermore, such as Figures 3 to 5 As shown, the inside of the cartridge 100 is also provided with a waste liquid chamber 170. The waste liquid chamber 170 is connected to the pyrolysis chamber 140 and the elution chamber 150 respectively through flow channels with valves. Specifically, the pyrolysis chamber 140 is connected to the waste liquid chamber 170 through the fifth flow channel 15 with the first valve 21, and the elution chamber 150 is connected to the waste liquid chamber 170 through the sixth flow channel 16 with the third valve 23.

[0064] from Figure 5 As can be seen, for flow channels with valves, such as the third flow channel 13, the portions located at both ends of the second valve 22 are respectively represented by bright solid lines and dark dashed lines, where the bright solid lines indicate that this portion of the flow channel is located at... Figure 5 On the front of the card box 100, the dark dotted lines indicate that this part of the flow channel is located... Figure 5 On the back of the card holder 100, which cannot be shown, the gray dotted lines indicate the transparent structure. The flow channels on both sides are interconnected through a through-hole penetrating the card holder 100, and both flow channels are sealed by a cover plate. This part... Figure 3 and Figure 4 As can be seen from the two perspectives shown, a valve is installed in the through-hole to control its opening or closing, thereby controlling the opening and closing of the corresponding flow channel.

[0065] Specifically, please refer to Figure 6 The figure shows Figure 4 The enlarged structure at point A mainly shows the explosion structure at the valve. As shown in the figure, a stepped surface 26 can be set in the first through hole 25, and a movable pressure plate 27 is set on the stepped surface 26 to form a valve. When the pressure plate 27 is separated from the stepped surface 26 or there is a gap, both ends of the first through hole 25 are in the open state, and the flow channels on both sides are interconnected. When the pressure plate 27 is pressed tightly on the stepped surface 26, the first through hole 25 is closed, and the flow channels on both sides are not interconnected.

[0066] The above is only one embodiment provided by the present application, which does not constitute a limitation on the specific structure of the present application. For example, in other embodiments, one flow channel can also be provided on one of the surfaces of the card box 100, or be formed in the interior of the card box 100, and the valve can be a blocking member provided in the flow channel. When the card box at the valve position is pressed, the wall of the flow channel is deformed and sealingly abuts against the valve, so that the flow channel is closed, and when the card box is released, the flow channel is opened.

[0067] Further, the card box 100 is provided with air holes respectively communicating with the lysis chamber 140 and the elution chamber 150. Please continue to refer to Figures 3 to 5 In the specific embodiment shown in the figure, the card box 100 is provided with a first air hole 31 and a second air hole 32, the first air hole 31 communicates with the lysis chamber 140, and the second air hole 32 communicates with the elution chamber 150. In other embodiments, only one air hole (i.e., the first air hole 31 and the second air hole 32 are the same air hole) can be provided on the card box 100, and the air hole is internally provided with two flow channels having valves, which respectively communicate with the lysis chamber 140 and the elution chamber 150, so that by controlling the valves on the corresponding flow channels, the air hole can be communicated with one of the lysis chamber 140 and the elution chamber 150 or both.

[0068] Please continue to refer to Figures 3 to 5 The card box 100 is also provided with reagent inlets respectively communicating with the lysis chamber 140 and the elution chamber 150. In Figures 3 to 5 the specific embodiment shown in the figure, the reagent inlets include a first inlet 41 and a second inlet 42 communicating with the lysis chamber 140, and a third inlet 43 and a fourth inlet 44 communicating with the elution chamber 150, wherein the first inlet 41 is used for the lysis reagent and the binding reagent to enter the lysis chamber 140, the second inlet is used for the washing reagent to enter the lysis chamber, the third inlet 43 is also used for the washing reagent to enter the elution chamber 150, and the fourth inlet 44 is used for the elution reagent to enter the elution chamber 150. Such a setting mode can arrange the corresponding reagent package at the actual inlet in advance during the preparation process of the detection, so as to save the operation of replacing the reagent package for injection during the detection process, and is beneficial to improving the detection efficiency. As with the air hole, the figure only provides one setting mode of the reagent inlet, and in other embodiments, only one reagent inlet can be provided, and two branch flow channels having valves are respectively communicated with the lysis chamber 140 and the elution chamber 150. When the reagent is injected into the corresponding chamber, the valve in the flow channel communicating with the chamber is controlled to be opened, and the other valve is closed. Of course, more reagent inlets can also be provided, which are not limited herein.

[0069] The above is the introduction and description of the specific structure of the card box 100. Based on the structure described above, the detection process of the card box 100 is specifically described below.

[0070] Before that, it should be noted that the detection process of the cartridge 100 can be operated manually or by a matching instrument. The following mainly describes the operation of the cartridge 100 in the product form shown in Figures 1-5 the instrument. Manual operation is to complete the actions of the instrument by manual operation. For some structural modifications compared to the illustrated embodiment, such as the arrangement of the valve, the number of air holes and reagent inlets, the essential detection principle is unchanged, and only the corresponding detailed operation method needs to be adjusted. See the above description of the valve, air hole and reagent inlet modification structure, and the following text will not be described in detail.

[0071] Please refer to Figure 4 and 5 , first, four reagent packs (the first reagent pack 51, the second reagent pack 52, the third reagent pack 53 and the fourth reagent pack 54 shown in Figure 4 ) containing corresponding reaction reagents are respectively placed at the first inlet 41, the second inlet 42, the third inlet 43 and the fourth inlet 44, and the corresponding reagent inlets are closed by the reagent packs themselves or additional sealing members.

[0072] Next, after adding the sample to the sample chamber 120 through the sample inlet 110, the sample inlet 110 is closed. Then the cartridge 100 is placed in the matching detection instrument, so that each valve abuts against the driving rod on the detection instrument, each air hole is connected with the driving pipeline of the detection instrument, and the driving rod of the detection instrument presses the pressing plate 27 on each valve to keep all valves in a closed state.

[0073] Next, negative pressure is introduced into the first air hole 31 through the driving pipeline, and the sample in the sample chamber 120 is drawn into the magnetic bead chamber 130, so that the sample is mixed with the magnetic beads in the magnetic bead chamber 130. In order to mix the sample and the magnetic beads uniformly, the positive / negative pressure can be introduced into the first air hole 31 reciprocally one or more times, so that the sample and the magnetic beads move reciprocally in the flow channel near the magnetic bead chamber 130, achieving the purpose of uniform mixing, or the magnetic beads and the sample can be treated by ultrasonic mixing. In addition to the negative pressure introduced into the first air hole 31, the second valve 22 can also be opened, and negative pressure can be introduced into the second air hole 32. The same applies to the other similar steps described below, which will not be described in detail.

[0074] Then continue to introduce negative pressure in the first air hole 31, the fluid formed after the sample and the magnetic beads are mixed is drawn into the lysis cavity 140. The first reagent bag 51 is squeezed to make the lysis agent and the binding agent in the first reagent bag 51 flow into the lysis cavity 140 through the first inlet 41, and the cells and / or viruses in the sample are lysed under the action of the lysis agent to expose the nucleic acid, and the exposed nucleic acid is combined with the magnetic beads under the action of the binding agent. Similarly, in order to make the lysis and combination fully react, the lysis cavity 140 can be ultrasonically mixed by the ultrasonic module of the detection instrument, of course, the mixing can also be performed by repeatedly shaking the cartridge or reciprocally introducing positive and negative pressure, and the mixing mode is not limited herein.

[0075] After the lysis and combination are completed, the magnetic beads are first adsorbed by the magnetic attraction module of the detection instrument to be fixed in the lysis cavity 140, then the first valve 21 is opened, and positive pressure is introduced in the first air hole 31 to drain the waste liquid in the lysis cavity 140 to the waste liquid cavity 170, after the liquid is drained, the pressure is stopped, the first valve 21 is closed, and the magnetic attraction module is removed to release the adsorption of the magnetic beads, at this time, the lysis cavity 140 contains the magnetic beads combined with the nucleic acid and some residual liquid and impurities.

[0076] The second reagent bag 52 is squeezed to make the washing agent in the second reagent bag 52 flow into the lysis cavity 140 through the second inlet 42 to wash the magnetic beads combined with the nucleic acid to remove other impurities adsorbed on the magnetic beads as much as possible, and the process can also be mixed to ensure that the washing is sufficient.

[0077] After the washing is completed, the second valve 22 is opened, and negative pressure is introduced in the second air hole 32 to draw the fluid in the lysis cavity 140 into the elution cavity 150, and then the second valve 22 is closed. Then the fluid drawn into the elution cavity 150 can be mixed again. It can be understood that in this step, the fluid in the lysis cavity 140 can also be pushed into the elution cavity 150 by introducing positive pressure in the first air hole 31, and the similar steps below are the same, and will not be described in detail.

[0078] Next, the magnetic beads are again adsorbed to be fixed in the elution cavity 150, then the third valve 23 is opened, and positive pressure is introduced in the second air hole 32 to make the waste liquid in the elution cavity 150 drain to the waste liquid cavity 170, after the liquid is drained, the positive pressure is stopped, the third valve 23 is closed, and the adsorption of the magnetic beads is released.

[0079] At this time, the magnetic beads may only be combined with the nucleic acid, or may still be combined with a small amount of impurities, in order to ensure the accuracy of the detection result, the magnetic beads can be washed again. Specifically, the third reagent bag 53 is squeezed to make the washing agent in the third reagent bag 53 flow into the elution cavity 150 through the third inlet 43 to wash the impurities adsorbed on the magnetic beads in the elution cavity 150, and the process can also be mixed to make the washing more sufficient.

[0080] After the cleaning, the magnetic beads in the elution cavity 150 are adsorbed and fixed, the third valve 23 is opened again, and a positive pressure is introduced through the second air hole 32 to discharge the waste liquid to the waste liquid cavity 170, then the third valve 23 is closed, the positive pressure is stopped, and the adsorption of the magnetic beads is released.

[0081] At this time, the magnetic beads are basically no longer adsorbed with impurities, but there are small liquid beads of residual detergent. Next, the magnetic beads are dried. Specifically, the second valve 22 is opened, and the elution cavity 150 is heated to make the small liquid beads on the magnetic beads in the elution cavity 150 vaporize. Since the second valve 22 is open, the vaporized and expanded small liquid beads flow into the lysis cavity 140 through the third flow channel 13. Since the lysis cavity 140 is at room temperature, the vaporized gas condenses on the inner wall of the lysis cavity 140 after entering the lysis cavity 140, so that it cannot flow back into the elution cavity 150. After drying is completed, the heating of the elution cavity 150 is stopped, and the second valve 22 is closed. It should be noted that, in addition to making the vaporized gas enter the lysis cavity 140 to condense by opening the second valve 22, the vaporized gas can also enter the waste liquid cavity 170 to condense by opening the third valve 23. Of course, the second valve 22 and the third valve 23 can also be opened at the same time.

[0082] After multiple cleaning and drying treatments, only the combined nucleic acid remains on the magnetic beads. Then, the fourth reagent bag 54 is squeezed to make the eluent in the fourth reagent bag 54 flow into the elution cavity 150 through the fourth inlet 44, so that the nucleic acid is eluted from the magnetic beads by the eluent. In order to ensure sufficient elution, mixing treatment can also be performed in this step.

[0083] After elution, the magnetic beads are first adsorbed and fixed in the elution cavity 150, then the fourth valve 24 is opened, and a positive pressure is introduced through the second air hole 32 to inject the liquid containing the eluted nucleic acid in the elution cavity 150 into the reaction cavity 160 through the fourth flow channel 14. In some embodiments, the reaction cavity 160 can pre-enclose a reaction reagent. The liquid containing the nucleic acid injected into the reaction cavity 160 is re-dissolved with the reaction reagent, and then the reaction cavity 160 is subjected to thermal cycling and optical analysis by a detection instrument, thereby completing all processes of nucleic acid detection. In order to simplify the production process of the cartridge 100, in other embodiments, the reaction cavity 160 can be a hollow cavity, and the reaction reagent can be injected from a reagent inlet connected with the lysis cavity 140 or the elution cavity 150, and then pressurized to enter the reaction cavity 160, or a reagent inlet connected with the reaction cavity 160 through a flow channel can be separately provided to inject the reaction reagent into the reaction cavity 160 through the reagent inlet.

[0084] It should be noted that the above detection process and the design position of the number of reagent inlets in the illustrated embodiments are only a specific way provided by the present application, and do not constitute a limitation on the specific number and position of the reagent inlets.

[0085] In order to ensure convenient sample injection, the sample inlet 110 is generally designed to be large, and the volume of the sample chamber 120 is also designed to be large. It is found through practice that the large volume of the sample chamber 120 will cause more sample liquid to be hung on the inner wall of the sample chamber 120 to form small liquid beads. Based on this, when the first gas hole 31 is in communication with the lysis chamber 140 in the manner shown in Figure 5 , in the process of introducing negative pressure into the first gas hole 31 to suck the sample liquid from the sample chamber 120 into the magnetic bead chamber 130 and the lysis chamber 140, due to the characteristics of negative pressure driving, the small liquid beads hung on the inner wall of the sample chamber 120 cannot be effectively sucked into the magnetic bead chamber 130 and the lysis chamber 140, so that this part of the small liquid beads cannot participate in the subsequent sample processing process, thereby affecting the processing capacity of the cartridge 100 to the sample.

[0086] In view of the above problems, in addition to the specific structure shown in Figures 1-6 , in some other embodiments, such as the two-sided perspective exploded structure and the perspective structure shown in Figures 7 to 10 , the first gas hole 31 can also be arranged to be in communication with the sample chamber 120. In the process of introducing positive pressure into the first gas hole 31 to pump the sample in the sample chamber 120 into the magnetic bead chamber 130, pump the fluid in the magnetic bead chamber 130 into the lysis chamber 140, and pump the fluid in the lysis chamber 140 into the waste liquid chamber 170, the direction of gas flow from the first gas hole 31 to the sample chamber 120 is as shown by the dashed arrow in Figure 9 between the first gas hole 31 and the sample chamber 120. Figure 5 and Figure 9 It can be seen that the specific embodiments shown in the two figures are basically the same in structure except that the chambers in communication with the first gas hole 31 are different.

[0087] After adopting the manner of the first gas hole 31 in communication with the sample chamber 120 and using the manner of introducing positive pressure into the first gas hole 31 to push the sample liquid to flow from the sample chamber 120 to the magnetic bead chamber 130 and the lysis chamber 140, based on the characteristics of positive pressure driving, the gas introduced into the sample chamber 120 will push most of the small liquid beads hung on the inner wall of the sample chamber 120 along the inner wall and the flow channel into the magnetic bead chamber 130 and the lysis chamber 140, so as to ensure that more sample liquid participates in the subsequent processing and detection, thereby improving the sample processing capacity and improving the detection effect.

[0088] Of course, in some other embodiments, at least two first air holes 31 can also be provided on the cartridge 100, at least one of which is in communication with the sample chamber 120, and at least one of which is in communication with the lysis chamber 140. When driving the sample, positive pressure can be introduced into the first air hole 31 in communication with the sample chamber 120, and negative pressure can be introduced into the first air hole 31 in communication with the lysis chamber 140.

[0089] In the cartridge 100, the magnetic bead chamber 130 can be independently provided in addition to the manner provided in the above embodiments that the magnetic bead chamber 130 is in communication with the sample chamber 120 and the lysis chamber 140 through the flow channel. Specifically, the sample chamber 120 is directly in communication with the lysis chamber 140 through the flow channel, the magnetic bead chamber 130 is independently provided in the cartridge 100, and the magnetic bead chamber 130 is in communication with the lysis chamber 140 through another flow channel. In addition, in order to enable the magnetic beads to smoothly enter the lysis chamber 140, the magnetic bead chamber 130 can contain a certain solution in addition to containing the magnetic beads, and an air hole in communication with the magnetic bead chamber 130 can be additionally provided on the cartridge 100. Based on this, in the detection process, the flow of the sample liquid from the sample chamber 120 to the lysis chamber 140 is basically the same as described above, and can be realized by introducing pressure into the first air hole 31, and the re-melting of the magnetic beads and the sample liquid can be realized by introducing positive pressure into the air hole on the cartridge 100 in communication with the magnetic bead chamber 130, so that the magnetic beads in the magnetic bead chamber 130 are driven into the lysis chamber 140 with the solution, and then the magnetic beads and the sample liquid are re-melted in the lysis chamber 140.

[0090] In summary, in the cartridge 100 provided in the embodiments of the present application, based on the research results that lysis and combination can be simultaneously performed and will not affect each other, only the lysis chamber 140 is provided in the cartridge 100 to be responsible for the lysis of nucleic acid and the combination with magnetic beads, and in order to prevent the impurities possibly remaining on the inner wall of the lysis chamber 140 from affecting the detection results, the elution chamber 150 is further provided to be responsible for the step of eluting the nucleic acid from the magnetic beads, so as to ensure the extraction purity of the nucleic acid in the eluted liquid, improve the accuracy of the detection results, and at the same time, as much as possible, reduce the volume of the cartridge 100 and the cost of the cartridge 100. Further, the waste liquid chamber 170 in communication with the lysis chamber 140 and the elution chamber 150 is provided, so as to ensure the orderly discharge of the waste liquid in the lysis chamber 140 and the elution chamber 150. Further, by using the valve provided on the flow channel between the chambers and the air hole in communication with part of the chambers, the detection process is simple to operate, and the internal fluid flow direction can be accurately controlled. At the same time, by using the temperature difference between the elution chamber 150 and the lysis chamber 140 and / or the waste liquid chamber 170 and controlling the valve therebetween, when the magnetic beads are dried, the vaporized gas can flow into the lysis chamber 140 and / or the waste liquid chamber 170 to be condensed, so as not to flow back into the elution chamber 150, and the purity of the nucleic acid extraction is further improved.

[0091] It needs to be particularly emphasized that a key factor to ensure the accuracy of nucleic acid detection is to avoid mixing the impurities generated by lysis into the solution after nucleic acid elution, because the solution after nucleic acid elution needs to be injected into the reaction chamber 160 for PCR amplification detection, and if the eluted solution is mixed with the impurities generated by lysis, the accuracy of PCR amplification detection will be affected. It is also for this reason that in the cartridge 100 provided in the embodiments of the present application, the lysis chamber 140 and the elution chamber 150 are independently arranged, the lysis chamber 140 is responsible for lysis, and the elution chamber 150 is responsible for elution. Compared with the same chamber for lysis and elution, this can prevent the impurities generated by lysis from affecting the elution, and through multiple washing, the impurities generated by lysis can be well avoided from mixing into the solution after elution.

[0092] In addition, the cartridge 100 provided in the embodiments of the present application also sets up independent air holes for lysis and elution, i.e. the first air hole 31 and the second air hole 32. The first air hole 31 is in communication with at least one of the sample chamber 120 and the lysis chamber 140, and is mainly responsible for driving the solution flowing between the sample chamber 120 and the lysis chamber 140, while the second air hole 32 is in communication with the elution chamber 150, and is responsible for driving the solution flowing from the elution chamber 150 to the waste chamber and the reaction chamber. Compared with the driving mode of setting only one air hole and driving the solution flowing by the same air hole, it can prevent the impurities remaining in the lysis chamber 140 from mixing into the elution chamber 150 during liquid driving, so as to avoid the impurities polluting the extraction product (i.e. the detection solution containing nucleic acid obtained after the magnetic beads are eluted) in the elution chamber 150, and prevent the subsequent nucleic acid detection result from being disturbed.

[0093] In order to improve the uniformity of the mixture of the magnetic beads and the sample, the present application further provides an embodiment, which is specifically described as follows: Figure 5 As shown in the figure, the second flow channel 12 includes a serpentine flow channel 121. The serpentine flow channel 121 can on the one hand increase the flow distance between the magnetic bead chamber 130 and the lysis chamber 140, so that the magnetic beads can be fully mixed with the sample during the flow process, and on the other hand, the curved corners thereof can also fully mix the magnetic beads in solid state with the sample in liquid state.

[0094] In addition, on the basis of setting the serpentine flow channel 121, by reciprocally introducing positive / negative pressure into the first air hole 31, the mixing effect of the magnetic beads and the sample can be optimized after the fluid formed by the magnetic beads and the sample reciprocally flows in the serpentine flow channel 121.

[0095] Considering that the injection of the liquid mixed with nucleic acid in the elution chamber 150 into the reaction chamber 160 will increase the pressure in the reaction chamber 160, and then affect the stability of the structure of the cartridge 100 at the reaction chamber 160, based on this, as shown in the figure, the cartridge 100 provided in the embodiments of the present application is provided with a third air hole 33, which is in communication with the reaction chamber 160 and the waste chamber 170, and is responsible for driving the solution flowing from the reaction chamber 160 to the waste chamber 170.Figure 5 As shown in the above embodiment, in some embodiments of the present application, the reaction cavity 160 is also communicated with the waste liquid cavity 170 through the seventh flow channel 17 with the water-proof air-permeable membrane, so that in the process of the liquid in the elution cavity 150 entering the reaction cavity 160, the gas in the reaction cavity 160 is discharged to the waste liquid cavity 170, and the liquid is blocked in the interior of the reaction cavity 160, while the water-proof air-permeable membrane can prevent the aerosol generated in the reaction cavity 160 from leaking to the outside to pollute the detection environment.

[0096] In Figure 4 and Figure 5 In the specific embodiment shown, similarly to the setting mode of the valve, the seventh flow channel 17 includes two parts respectively opened on the two surfaces of the cartridge 100, and the two parts are communicated through the second through hole 162 penetrating the cartridge 100, and the water-proof air-permeable membrane 161 is covered on the second through hole 162 to allow the gas to pass through and the liquid to be blocked. Of course, in some other embodiments, the seventh flow channel 17 can be formed only on one surface of the cartridge 100 or be injection molded in the interior of the cartridge 100, and the water-proof air-permeable membrane 161 is arranged to cover the entire cross section of the pipeline of the seventh flow channel 17.

[0097] By communicating the reaction cavity 160 with the waste liquid cavity 170 through the seventh flow channel 17 and arranging the water-proof air-permeable membrane 161 in the seventh flow channel 17, when the liquid is injected into the reaction cavity 160, the gas in the reaction cavity 160 can be normally discharged into the waste liquid cavity 170, so as to ensure the stability of the pressure in the reaction cavity 160.

[0098] In addition to the mode of ensuring the stability of the pressure in the reaction cavity 160 by arranging the seventh flow channel 17 and the water-proof air-permeable membrane 161, the reaction cavity 160 can also be subjected to vacuumizing treatment in the manufacturing process of the cartridge 100, so as to ensure the smoothness of the liquid injection into the reaction cavity 160 and the stability of the pressure in the interior of the reaction cavity 160. Alternatively, a material with good structural strength and strong pressure resistance can also be used to prepare the cartridge 100, so as to ensure that the reaction cavity 160 still has good stability when the pressure in the reaction cavity 160 increases.

[0099] In order to improve the accuracy of the detection, the present application further provides an embodiment, and the specific reference is made to Figure 5 As shown in the figure, the fourth flow channel 14 and the seventh flow channel 17 are both provided with the blocking valve 163, which is configured to be closed after the liquid injection into the reaction cavity 160 is completed.

[0100] Specifically, the blocking valve 163 can adopt the tabletting form provided in the above structural description about the valve, or can adopt Figure 5The thin film valve provided in the embodiment shown, specifically, blocks the hole on the corresponding flow channel by pressing the thin film to block the two sections of the corresponding flow channel. After the liquid injection in the reaction cavity 160 is completed, the blocking valve 163 is closed to seal the reaction cavity 160, so that the reaction cavity 160 can maintain a certain volume for thermal cycling reaction when the reaction cavity is subjected to thermal cycling and detection, thereby ensuring the accuracy of the detection result.

[0101] In order to reduce the detection error and ensure the accuracy of the detection result, in some embodiments, as shown in Figure 5 , the reaction cavity 160 is multiple, and the multiple reaction cavities 160 are arranged in parallel. Specifically, the multiple reaction cavities 160 arranged in parallel means that each reaction cavity 160 is independently communicated with other cavities through the corresponding flow channel and valve. During the liquid injection into the reaction cavity 160, the liquid with nucleic acid in the elution cavity 150 will fill each reaction cavity 160 respectively, and during the subsequent detection, the detection liquid in each reaction cavity 160 will be detected respectively to meet the detection needs of different projects and improve the sample detection capability.

[0102] In order to improve the detection efficiency of the detection liquid in the reaction cavity 160, the present application further provides an embodiment, please refer to Figure 11 , the bottom structure of the cartridge 100 is shown, as shown in the figure, one side of the cartridge 100 is formed with a sheet structure 180, the reaction cavity 160 is arranged in the inside of the sheet structure 180, and the wall thickness of the sheet structure 180 is smaller than that of the rest of the cartridge 100.

[0103] In the embodiment, by arranging the reaction cavity 160 in the sheet structure 180 with thinner wall thickness, when the detection liquid in the reaction cavity 160 is subjected to PCR detection, the heat loss when the heat passes through the wall of the reaction cavity 160 can be reduced, thereby improving the heat conduction efficiency, shortening the amplification time, and finally achieving the purpose of improving the detection efficiency.

[0104] For reagent injection, in addition to placing the existing reagent package at the reagent inlet of the cartridge 100 for liquid injection, the reagent package can also be integrated directly on the cartridge 100. Please refer to Figure 1 and Figure 4 , as shown in the figure, the reagent inlet is sealed and covered with a reagent package (the first reagent package 51, the second reagent package 52, the third reagent package 53 and the fourth reagent package 54 in the figure, the specific number of reagent packages is not limited), and the reagent package contains reagents.

[0105] The reagent bag can be a flexible vesicle and realized by film hot pressing or the like. The specific implementation is not limited here. After the card box 100 and the reagent bag are produced separately, the reagent bag is assembled and fixed at the reagent inlet of the card box 100 by means of gluing, hot welding or the like, and the reagent inlet is sealed to prevent the aerosol in the internal space of the card box 100 and the reagent after vaporization from leaking through the reagent inlet to cause environmental pollution.

[0106] Further, in order to facilitate the reagent bag to be broken for liquid injection, in some embodiments, a piercing structure is arranged at the reagent inlet, and the reagent bag is used to be pressed and pierced by the piercing structure when stressed, so that the reagent inside enters the internal space of the card box 100 through the reagent inlet.

[0107] Specifically, the piercing structure can be a conical protrusion, a needle or the like. By arranging the piercing structure, on the one hand, the corresponding reagent bag can be easily pierced when it is needed to inject the reagent, and on the other hand, the position of the piercing structure can realize the control of the piercing position of the reagent bag, so as to avoid the reagent bag from being broken seriously due to excessive pressing, and thus the seal between the reagent inlet and the external environment is damaged, causing environmental pollution.

[0108] Considering that the pressure in the waste liquid chamber 170 will be increased after the liquid is discharged into the waste liquid chamber 170, which may affect the structural stability of the card box 100, based on this, the present application further proposes an embodiment, please refer to Figure 3 and Figure 5 As shown in the figure, the card box 100 is also provided with a pressure relief hole 60 which communicates with the waste liquid chamber 170, and the pressure relief hole 60 is used to release the gas in the waste liquid chamber 170 during the process of discharging liquid into the waste liquid chamber 170, so as to ensure the stability of the pressure in the waste liquid chamber 170.

[0109] Similarly to the release of the gas in the reaction chamber 160 during the liquid injection mentioned in the above embodiments, in addition to the pressure relief hole 60, the waste liquid chamber 170 can also be evacuated or made of a material with good structural strength and high pressure resistance to ensure the stability of the structure of the waste liquid chamber 170.

[0110] In order to fully reduce the pollution to the detection environment, the present application further proposes an embodiment, specifically, at least one of the sample chamber 120 and the lysis chamber 130 and the first gas hole 31, the elution chamber 150 and the second gas hole 32, and the pressure relief hole 60 and the waste liquid chamber 170 are covered with a waterproof and breathable film, which is used to prevent the aerosol and the reagent after vaporization in the internal space of the card box 100 from leaking. In Figure 3In the specific embodiment shown, the first gas hole 31 and the lysis chamber 140 are covered by a first waterproof and air-permeable film 71, the second gas hole 32 and the elution chamber 150 are covered by a second waterproof and air-permeable film 72, and the pressure relief hole 60 and the waste liquid chamber 170 are covered by a third waterproof and air-permeable film 73. In Figure 8 In the specific embodiment shown, the first waterproof and air-permeable film 71 is located between the first gas hole 31 and the sample chamber 120, and the second waterproof and air-permeable film 72 and the third waterproof and air-permeable film 73 are located in the same positions as in the embodiment shown. Figure 3

[0111] Specifically, the first waterproof and air-permeable film 71 can be covered on the opening of the first gas hole 31 as shown in Figure 3 and Figure 8 , or can be covered in the flow channel between the first gas hole 31 and the lysis chamber 140 or the sample chamber 120, and the second waterproof and air-permeable film 72 and the third waterproof and air-permeable film 73 are the same, which will not be described here.

[0112] In this embodiment, by closing the channels such as gas holes and pressure relief holes 60 that connect the internal space of the card box 100 and the external space with waterproof and air-permeable films, internal aerosol pollution of the detection environment can be effectively prevented.

[0113] In some embodiments, at least one of the sample chamber 120 and the lysis chamber 140 and the first gas hole 31, and the elution chamber 150 and the second gas hole 32 are provided with a water-absorbing and air-permeable film, which is used to prevent water from entering the internal space of the card box 100 when the gas hole introduces positive pressure, affecting the accuracy of the detection result, and the bubbles will break when they hit the water-absorbing and air-permeable film, thereby eliminating the bubbles. In addition, the water-absorbing and air-permeable film also has the function of absorbing the water vapor evaporated during the drying of the magnetic beads. Figure 4 and Figure 5 In the specific embodiment shown, the first gas hole 31 and the lysis chamber 140 are provided with a first water-absorbing and air-permeable film 81, and the second gas hole 32 and the elution chamber 150 are provided with a second water-absorbing and air-permeable film 82. In Figure 8 and Figure 9 In the specific embodiment shown, the first water-absorbing and air-permeable film 81 is provided between the first gas hole 31 and the sample chamber 120, and the second water-absorbing and air-permeable film 82 is provided in the same position as in Figure 4 and Figure 5 the embodiment shown.

[0114] Considering that the air permeability of some water-absorbing and air-permeable films may decrease after absorbing a large amount of liquid, the liquid inside the chamber will splash during the ultrasonic vibration mixing of the lysis chamber 140 and the elution chamber 150, based on which, as shown in Figure 4 and Figure 5 ​As shown, a significant amount of liquid may pass through the first channel 91 between the lysis chamber 140 and the first absorbent gas membrane 81 and be absorbed by the first absorbent gas membrane 81. Similarly, a significant amount of liquid may pass through the second channel 92 between the elution chamber 150 and the second absorbent gas membrane 82 and be absorbed by the second absorbent gas membrane 82. This results in a decrease in the permeability of the first and second absorbent gas membranes 81 and 82. Consequently, when gas pressure is introduced through the first pore 31 and the second pore 32, the permeability of the gas at the first and second absorbent gas membranes 81 and 82 decreases, reducing the driving ability of the internal liquid. In severe cases, this may even prevent the sample from flowing normally between the internal chambers and channels.

[0115] contrast Figure 9 and Figure 5 It can be seen that, Figure 9 The illustrated embodiment is compared to Figure 5 In the illustrated embodiment, since the first absorbent and breathable membrane 81 is located between the first pore 31 and the sample chamber 120, during ultrasonic mixing in the lysis chamber 140, the long and tortuous flow channel between the first absorbent and breathable membrane 81 and the lysis chamber 140 can prevent splashed liquid from reaching the first absorbent and breathable membrane 81 and being absorbed by it. This ensures the breathability of the first absorbent and breathable membrane 81 and guarantees the driving effect of positive pressure on the liquid when the first pore 31 is introduced. Furthermore, even if some liquid splashes into the flow channel between the lysis chamber 140 and the sample 120 or into the sample chamber 120, positive pressure can be introduced through the first pore 31 to push this portion back into the lysis chamber 140 to participate in subsequent reactions, ensuring the sample processing capacity.

[0116] As for the second absorbent and breathable membrane 82 between the second pore 32 and the elution chamber 150, such as Figure 9 As shown, due to Figure 10 The second waterproof and breathable membrane 72 at the second pore 32 shown is prone to failure when encountering large air bubbles. Therefore, a micro valve 921 is added to the second channel 92 between the second absorbent breathable membrane 82 and the elution chamber 150. The cross-sectional area of ​​the second channel 92 at the micro valve 921 is reduced and covered with an elastic pad. The function of the micro valve 921 is to eliminate or transform large air bubbles that splash into the second channel 92 when the sample in the elution chamber 150 is ultrasonically mixed, under the impact of the elastic pad and when entering the smaller gap section from a larger cross section. This allows the large air bubbles to be quickly absorbed when they reach the second absorbent breathable membrane 82, preventing large air bubbles from passing through the second channel 92 and penetrating the second absorbent breathable membrane 82 and reaching the second waterproof and breathable membrane 72, causing the second waterproof and breathable membrane 72 to fail.

[0117] Of course, the micro valve 921 can also be set as a flow blocking valve. Before the sample in the elution cavity 150 is ultrasonically mixed, the micro valve 921 can be closed in advance to ensure that the liquid cannot splash to the second water-absorbing and air-permeable film 82 and affect the air permeability of the second water-absorbing and air-permeable film 82, and ensure the driving ability of the sample when the pressure is introduced into the second air hole 32 later.

[0118] In addition, Figure 9 and 10 In the embodiment shown, the first water-absorbing and air-permeable film 81 and the second water-absorbing and air-permeable film 82 are arranged at the edges or corners of the cartridge 100. This is because the middle area of the cartridge 100 needs to integrate more cavities and flow channels. In order to ensure that the first water-absorbing and air-permeable film 81 and the second water-absorbing and air-permeable film 82 have a larger area, they can absorb more liquid without being blocked. Therefore, the first water-absorbing and air-permeable film 81 and the second water-absorbing and air-permeable film 82 are arranged at the edges or corners of the cartridge 100, so that the area of the first water-absorbing and air-permeable film 81 and the second water-absorbing and air-permeable film 82 can be as large as possible.

[0119] It is considered that the way of integrally forming micro flow channels in the cartridge 100 has the problems of complex process and high cost. Therefore, the application provides a manufacturing method which is convenient for batch generation of the cartridge 100 and can effectively reduce the cost. For details, please refer to Figures 1 to 4 As shown in the figure, the cartridge 100 includes a cartridge main body 101 and a cover 102. The sample inlet 110, the air hole and the reagent inlet are all arranged on the cartridge main body 101. The sample cavity 120, the magnetic bead cavity 130, the lysis cavity 140, the elution cavity 150, the reaction cavity 160, the waste liquid cavity 170 and the flow channels connecting them are all formed by grooves arranged on the surface of the cartridge main body 101, and the cover 102 seals and covers these grooves.

[0120] In the specific embodiment shown in the figure, grooves forming flow channels are arranged on both surfaces of the cartridge main body 101. Correspondingly, the cover 102 covers the grooves on both surfaces of the cartridge main body 101. In the specific production process, in order to ensure the structural strength, the cartridge main body 101 can be made of hard material, and in order to ensure the sealing performance, the cover 102 can be made of flexible film material. The two can be assembled and fixed by adhesion, heat sealing or the like.

[0121] Compared with the way of integrally forming flow channels and cavities inside, the way of arranging grooves on the cartridge main body 101 and sealing the grooves by the cover 102 provided in the embodiment is more convenient for the production and manufacturing of the cartridge 100, which is beneficial to improve the production efficiency and reduce the production cost.

[0122] Please refer to Figure 4In some embodiments of the present application, a water absorption member 171 is arranged in the waste liquid cavity 170, and is used to absorb the liquid discharged into the waste liquid cavity 170, so as to prevent the waste liquid from leaking and facilitate subsequent treatment of the waste liquid.

[0123] In order to ensure the smoothness of the sample flow, as shown in Figure 4 and Figure 5 In some embodiments, the first flow channel 11 is a siphon channel, so that in the process of drawing the sample in the sample cavity 120 into the magnetic bead cavity 130, the sample flow can be ensured to be sufficiently smooth by using the siphon principle.

[0124] In order to facilitate the sealing of the sample inlet 110 after sample injection, as shown in Figure 1 , the detachable cover at the sample inlet 110 is provided with a sealing cover 111. Specifically, the sealing cover 111 can seal the sample inlet 110 by threaded connection, extrusion interference fit and the like.

[0125] Based on the cartridge 100 provided in the above embodiments, according to another aspect of the embodiments of the present application, there is further provided a nucleic acid detection method, which is applied to the cartridge 100 in any of the above embodiments. For details, please refer to Figure 5 , and further in combination with Figure 12 , the flow of the nucleic acid detection method is shown in the figure, as shown in the figure, the method comprises the following steps:

[0126] Step 210: injecting the sample into the sample cavity 120 through the sample inlet 110;

[0127] Step 230: keeping all the valves closed;

[0128] Step 251: introducing negative pressure into the first air hole 31, so that the sample is drawn into the magnetic bead cavity 130 to mix with the magnetic beads to form a first mixed fluid;

[0129] Step 271: introducing negative pressure into the first air hole 31 again, so that the first mixed fluid is drawn into the lysis cavity 140;

[0130] When the method is applied to the cartridge 100 shown in Figures 7 to 10 , steps 251 and 271 are changed to Figure 13In steps 252 and 272, the positive pressure introduced into the first vent 31 has the same driving direction and effect on the sample and the first mixed fluid as in steps 251 and 271. However, when this method is applied to an embodiment where multiple first vents 31 are connected to the sample chamber 120 and the lysis chamber 140 respectively, steps 251 and 271 are changed to introduce positive pressure into the first vent 31 connected to the sample chamber 120 and negative pressure into the first vent 31 connected to the lysis chamber 140. The driving direction and effect on the sample and the first mixed fluid are still the same. Similar steps in the following text are similar and will not be described in detail.

[0131] Step 290: Through the reagent inlet connected to the lysis chamber 140 ( Figure 5 The first inlet 41) injects lysing agent and binding agent into the lysis chamber 140, so that the lysing agent and binding agent are mixed with the first mixed fluid and react to form a second mixed fluid. In the first mixed fluid, the cells and / or viruses in the lysing agent are lysed to expose the nucleic acid, and the nucleic acid is bound to the magnetic beads under the action of the binding agent.

[0132] Step 310: Adsorb the magnetic beads in the second mixed fluid to fix the magnetic beads with bound nucleic acids in the lysis chamber 140, and open the flow channel between the lysis chamber 140 and the waste liquid chamber 170. Figure 5 The first valve 21 on the fifth flow channel 15) introduces positive pressure into the first vent 31, so that the liquid in the pyrolysis chamber 140 is discharged to the waste liquid chamber 170. Then the first valve 21 is closed and the magnetic bead is released.

[0133] Step 330: Through the reagent inlet connected to the lysis chamber 140 ( Figure 5 The detergent is injected into the pyrolysis chamber 140 through the second inlet 42) to clean the magnetic beads and form a third mixed fluid;

[0134] Step 351: Open the flow channel between the pyrolysis chamber 140 and the elution chamber 150 ( Figure 5 The second valve 22 on the third flow channel 13) is used to introduce negative pressure into the second vent 32, which communicates with the elution chamber 150, so that the third mixed fluid is drawn into the elution chamber 150, and then the second valve 22 is closed. Of course, this step can also be replaced by... Figure 13 Step 352 shown in the diagram achieves the same driving effect on the third mixed fluid by introducing positive pressure through the first vent 31. Similar steps in the following text are similar and will not be described in detail.

[0135] Step 370: Adsorb the magnetic beads in the third mixed fluid to fix the magnetic beads in the elution chamber 150, and open the flow channel between the elution chamber 150 and the waste liquid chamber 170. Figure 5Step 430: open the second valve 22 and / or the third valve 23, and dry the magnetic beads in the elution chamber 150, so that the liquid on the surface of the magnetic beads and the liquid in the elution chamber 150 are vaporized and then condensed in the lysis chamber 140 and / or the waste liquid chamber 170, and then close the second valve 22 and / or the third valve 23.

[0136] Step 450: inject an eluent into the elution chamber 150 through the reagent inlet (the fourth inlet 44) connected to the elution chamber 150, so as to elute the nucleic acid from the magnetic beads. Figure 5

[0137] Step 410: adsorb the magnetic beads in the fourth mixed fluid, so that the magnetic beads are fixed in the elution chamber 150, open the third valve 23, and introduce positive pressure into the second air hole 32, so that the liquid in the elution chamber 150 is discharged to the waste liquid chamber 170, and then close the third valve 23 and release the adsorption of the magnetic beads.

[0138] Step 430: open the second valve 22 and / or the third valve 23, and dry the magnetic beads in the elution chamber 150, so that the liquid on the surface of the magnetic beads and the liquid in the elution chamber 150 are vaporized and then condensed in the lysis chamber 140 and / or the waste liquid chamber 170, and then close the second valve 22 and / or the third valve 23.

[0139] Step 450: inject an eluent into the elution chamber 150 through the reagent inlet (the fourth inlet 44) connected to the elution chamber 150, so as to elute the nucleic acid from the magnetic beads. Figure 5 Step 470: adsorb the magnetic beads in the elution chamber 150, so that the magnetic beads are fixed in the elution chamber 150, open the fourth valve 24 on the flow channel (the fourth flow channel 14) connected the elution chamber 150 and the reaction chamber 160, and introduce positive pressure into the second air hole 32, so as to inject the liquid mixed with the nucleic acid in the elution chamber 150 into the reaction chamber 160 for detection.

[0140] Figure 5 It should be noted that if the lysis agent or other impurity solution remains in the elution chamber 150, it will contaminate the subsequent injected eluent, so that the liquid mixed with the nucleic acid and injected from the elution chamber 150 into the reaction chamber 160 contains impurities, which affects the PCR amplification detection.

[0141] It should be noted that if the lysis agent or other impurity solution remains in the elution chamber 150, it will contaminate the subsequent injected eluent, so that the liquid mixed with the nucleic acid and injected from the elution chamber 150 into the reaction chamber 160 contains impurities, which affects the PCR amplification detection.

[0142] ​​Therefore, in the nucleic acid detection method provided by the embodiment of the present application, after the step 310 preliminarily discharges the lysis agent and impurities into the waste liquid chamber 170, the step 330 is used to inject a detergent into the lysis chamber 140 to clean the residual lysis agent and other impurities in the lysis chamber 140 to form a third mixed fluid, the step 351 is used to drive the third mixed fluid into the elution chamber 150, and the step 370 is used to discharge the impurities (including the residual lysis agent, the detergent, etc.) in the third mixed fluid from the elution chamber 150 to the waste liquid chamber 170, so as to complete the first washing and liquid discharge. Then, the step 390 is used to inject the detergent into the elution chamber 150 again to clean the residual impurities, and the step 390 is used to discharge the impurities (including the detergent and the residual lysis agent, etc.) in the elution chamber 150 to the waste liquid chamber 170, so as to complete the second washing and liquid discharge. In this way, the impurities in the elution chamber 150 are removed by twice washing, and the magnetic beads in the elution chamber 150 are dried by the step 450, so that the liquid beads remaining in the elution chamber 150 are evaporated and discharged, the purity of the solution in the elution chamber 150 after the step 450 is used to inject the elution liquid into the elution chamber 150 is ensured, and the accuracy of the nucleic acid detection in the reaction chamber in the step 470 is improved.

[0143] Specifically, in the step 470, the reaction reagent can be pre-contained in the reaction chamber 160. After the liquid mixed with the nucleic acid injected from the elution chamber 150 reacts with the reaction reagent, the detection liquid used for PCR detection is formed. Of course, the reaction reagent can also be injected into the reaction chamber through the reagent inlet in communication with the elution chamber 150 or the lysis chamber 140.

[0144] The flow of the above nucleic acid detection method is the same as the description of the nucleic acid detection process in the nucleic acid detection cartridge embodiment provided above, and will not be described again here.

[0145] The nucleic acid detection method provided by the embodiment of the present application can ensure the extraction purity of the nucleic acid in the eluted liquid, improve the accuracy of the detection result, and has simple operation in the whole detection process, and can realize accurate control of the fluid in the cartridge 100.

[0146] Further, for the cartridge 100 in which the magnetic bead chamber 130 and the lysis chamber 140 are in communication through the serpentine flow channel 121, the step 251 in the nucleic acid detection method can include:

[0147] The first air hole 31 in communication with the lysis chamber 140 is reciprocally introduced into positive / negative pressure n times, so that the sample and the magnetic beads reciprocally flow and mix in the magnetic bead chamber 130 and the serpentine flow channel 121 to form a first mixed fluid, wherein n≥1, and n is an integer.

[0148] To ensure sufficient reaction in each step, the fluid is mixed after step 290, and / or after step 330, and / or after step 351, and / or after step 390, and / or after step 450. Mixing methods include, but are not limited to, ultrasonic mixing, shaking mixing, etc.

[0149] When this method is applied Figures 7 to 10 When the cartridge 100 is shown, before introducing pressure into the second vent 32, the microvalve 921 can be opened to ensure that the airflow enters the elution chamber 150 normally and drives the internal fluid flow. Before mixing the internal fluid, such as before ultrasonic mixing, the microvalve 921 can be closed to ensure that splashed fluid does not reach the second absorbent membrane 82 and be absorbed by it.

[0150] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of this application, and not to limit them. Although this application has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some or all of the technical features therein; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of this application. In particular, as long as there is no structural conflict, the various technical features mentioned in the various embodiments can be combined in any way.

Claims

1. A nucleic acid testing cartridge, characterized by, The cartridge is provided with a sample inlet, and a sample chamber is arranged in the cartridge and communicates with the sample inlet; The cartridge is further provided with a lysis chamber, an elution chamber and a reaction chamber which are sequentially communicated through flow channels with valves, and the sample chamber communicates with the lysis chamber through a flow channel; The cartridge is further provided with a magnetic bead chamber, and the magnetic bead chamber contains magnetic beads, and the magnetic bead chamber communicates with the lysis chamber through a flow channel; The cartridge is further provided with a waste liquid chamber, and the waste liquid chamber communicates with the lysis chamber and the elution chamber through flow channels with valves, respectively; The cartridge is provided with a first air hole which communicates with at least one of the sample chamber and the lysis chamber; The cartridge is further provided with a second air hole which communicates with the elution chamber; The cartridge is further provided with a reagent inlet which respectively communicates with the lysis chamber and the elution chamber.

2. The nucleic acid testing cartridge of claim 1, wherein, The magnetic bead chamber is communicated between the sample chamber and the lysis chamber through a flow channel, so that the sample chamber communicates with the lysis chamber through a flow channel and the magnetic bead chamber.

3. The nucleic acid testing cartridge of claim 2, wherein, The first air hole communicates with the sample chamber, and the first air hole is used to drive the sample from the sample chamber into the magnetic bead chamber and from the magnetic bead chamber into the lysis chamber when a positive pressure is introduced.

4. The nucleic acid testing cartridge of claim 1, wherein, The reaction chamber further communicates with the waste liquid chamber through a flow channel with a water-proof and air-permeable membrane, so that in the process of liquid in the elution chamber entering the reaction chamber, the gas in the reaction chamber is discharged to the waste liquid chamber, and the liquid is blocked in the interior of the reaction chamber, while preventing the leakage of aerosol in the reaction chamber.

5. The nucleic acid testing cartridge of claim 4, wherein, A blocking valve is arranged in the flow channel between the reaction chamber and the elution chamber and the flow channel between the reaction chamber and the waste liquid chamber, and the blocking valve is configured to be closed after the liquid injection in the reaction chamber is completed.

6. The nucleic acid testing cartridge of claim 1, wherein, The reaction chamber contains a reaction reagent, and the reaction reagent is used to react with the liquid injected into the reaction chamber from the elution chamber.

7. The nucleic acid testing cartridge of claim 1, wherein, One side of the cartridge is formed with a sheet structure, the reaction chamber is arranged in the interior of the sheet structure, and the wall thickness of the sheet structure is smaller than that of the remaining part of the cartridge.

8. The nucleic acid testing cartridge of claim 1, wherein, The reaction chamber is a plurality of chambers, and the plurality of chambers are arranged in parallel.

9. The nucleic acid testing cartridge of claim 1, wherein, The reagent inlet includes a first inlet and a second inlet which respectively communicate with the lysis chamber, the first inlet is used for introducing a lysis reagent and a binding reagent into the lysis chamber, and the second inlet is used for introducing a washing reagent into the lysis chamber; The reagent inlet further includes a third inlet and a fourth inlet, the third inlet is used for introducing a washing reagent into the elution chamber, and the fourth inlet is used for introducing an elution reagent into the elution chamber.

10. The nucleic acid testing cartridge of claim 1, wherein, The cartridge is further provided with a pressure relief hole which communicates with the waste liquid chamber.

11. The nucleic acid testing cartridge of claim 10, wherein, At least one of the sample chamber and the lysis chamber, the elution chamber, the pressure relief hole and the waste liquid chamber are covered with a water-proof and air-permeable membrane, and the water-proof and air-permeable membrane is used to prevent the leakage of aerosol and vaporized reagent in the interior.

12. A method of detecting a nucleic acid, characterized by, The nucleic acid detection cartridge of any one of claims 2-11 is applied to the method, and the method comprises: injecting a sample into the sample chamber through the sample inlet; keeping all valves closed; introducing pressure into the first air hole to make the sample flow into the magnetic bead cavity and mix with the magnetic beads to form a first mixed fluid; introducing pressure into the first air hole again to make the first mixed fluid flow into the lysis cavity; injecting a lysis agent and a binding agent into the lysis cavity through a reagent inlet connected with the lysis cavity to make the lysis agent and the binding agent mix and react with the first mixed fluid to form a second mixed fluid, wherein the cells and / or viruses in the first mixed fluid are lysed by the lysis agent to expose nucleic acid, and the nucleic acid is bound to the magnetic beads by the binding agent; adsorbing the magnetic beads in the second mixed fluid to fix the magnetic beads with the nucleic acid in the lysis cavity, opening a first valve on a flow channel between the lysis cavity and the waste liquid cavity, introducing positive pressure into the first air hole to make the liquid in the lysis cavity flow into the waste liquid cavity, then closing the first valve and releasing the adsorption of the magnetic beads; injecting a washing agent into the lysis cavity through the reagent inlet connected with the lysis cavity to clean the magnetic beads and form a third mixed fluid; opening a second valve on a flow channel between the lysis cavity and the elution cavity, introducing positive pressure into the first air hole or negative pressure into the second air hole to make the third mixed fluid flow into the elution cavity, then closing the second valve; adsorbing the magnetic beads in the third mixed fluid to fix the magnetic beads in the elution cavity, opening a third valve on a flow channel between the elution cavity and the waste liquid cavity, introducing positive pressure into the first air hole or the second air hole to make the liquid in the elution cavity flow into the waste liquid cavity, then closing the third valve and releasing the adsorption of the magnetic beads; injecting a washing agent into the elution cavity through the reagent inlet connected with the elution cavity to clean the magnetic beads and form a fourth mixed fluid; adsorbing the magnetic beads in the fourth mixed fluid to fix the magnetic beads in the elution cavity, opening the third valve, introducing positive pressure into the first air hole or the second air hole to make the liquid in the elution cavity flow into the waste liquid cavity, then closing the third valve and releasing the adsorption of the magnetic beads; opening the second valve and / or the third valve and drying the magnetic beads in the elution cavity to make the liquid on the surface of the magnetic beads and the liquid in the elution cavity vaporize and then condense in the lysis cavity and / or the waste liquid cavity, then closing the second valve and / or the third valve; injecting an elution agent into the elution cavity through the reagent inlet connected with the elution cavity to elute the nucleic acid from the magnetic beads; adsorbing the magnetic beads in the elution cavity to fix the magnetic beads in the elution cavity, opening a fourth valve on a flow channel between the elution cavity and the reaction cavity, and introducing positive pressure into the first air hole or the second air hole to inject the liquid mixed with the nucleic acid in the elution cavity into the reaction cavity for detection.

13. The nucleic acid detection method according to claim 12, wherein After the step of injecting a lysis reagent and a binding reagent into the lysis chamber through a reagent inlet in communication with the lysis chamber, mixing and reacting the lysis reagent and the binding reagent with the first mixed fluid to form a second mixed fluid, and / or, after the step of injecting a washing reagent into the lysis chamber through a reagent inlet in communication with the lysis chamber to wash the magnetic beads and form a third mixed fluid, and / or, after the step of opening a second valve on a flow path between the lysis chamber and the elution chamber, and introducing a positive pressure through the first air hole or a negative pressure through the second air hole to allow the third mixed fluid to flow to the elution chamber, and then closing the second valve, and / or, after the step of injecting a washing reagent into the elution chamber through a reagent inlet in communication with the elution chamber to wash the magnetic beads and form a fourth mixed fluid, and / or, after the step of injecting an elution reagent into the elution chamber through a reagent inlet in communication with the elution chamber to elute the nucleic acid from the magnetic beads, the method further comprises the step of: mixing the fluid.

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