Nucleic acid detection card box
By designing a nucleic acid detection box containing multiple functional chambers and microflowers, the problems of complex nucleic acid detection operations and high contamination risks are solved, and a simple and efficient nucleic acid extraction, amplification and hybridization process is achieved.
Patent Information
- Application Number
- CN202421438866.6
- Authority / Receiving Office
- CN · China
- Patent Type
- Utility models(China)
- Current Assignee / Owner
- Filing Date
- 2024-06-21
- Publication Date
- 2025-04-15
- Estimated Expiration
- 2034-06-21
AI Technical Summary
Nucleic acid testing operations are complex, cumbersome and time-consuming, and the risk of failure caused by the introduction of pollution is high.
A nucleic acid detection card box is provided, including a substrate, a sample filling chamber, a liquid sac, a mixing chamber, a pressurized chamber, a column mold cavity, an amplification chamber and a hybrid cavity. The extraction, amplification and hybridization operations of samples are achieved through microflower connections, reducing manual operation and liquid transfer.
The nucleic acid detection operations are simplified, the detection efficiency is improved, the risk of pollution is reduced, and the automation and efficiency of nucleic acid extraction, amplification and hybridization are realized.
Smart Images
Figure CN222758234U_ABST
Abstract
Description
Technical Field
[0001] The present application relates to the field of nucleic acid detection technology, and in particular to a nucleic acid detection card box. Background Art
[0002] At present, when conducting nucleic acid testing, extraction, amplification and hybridization operations need to be completed. The entire operation process needs to be completed between multiple laboratories through manual control of different equipment. Manual transfer is also required in the middle. The operation is complicated, tedious and time-consuming, and there is a high risk of failure due to introduction of contamination. Utility Model Content
[0003] The main purpose of the present application is to provide a nucleic acid detection card box, which aims to solve the problem that the nucleic acid detection operation is complicated, tedious and time-consuming, and the risk of failure due to the introduction of contamination is high.
[0004] To achieve the above-mentioned purpose, the present application provides a nucleic acid detection card box, which includes a substrate, a sample filling chamber, a plurality of liquid capsules, a mixing chamber, a first pressurizing chamber, a column mold cavity, an amplification chamber and a hybridization chamber. The substrate has a first seat surface and a second seat surface that are arranged opposite to each other in the thickness direction thereof, and a plurality of liquid capsule grooves are arranged on the side of the first seat surface away from the second seat surface; the sample filling chamber is arranged on the side of the first seat surface away from the second seat surface; the plurality of liquid capsules are all arranged on the side of the first seat surface away from the second seat surface and correspond one-to-one to the plurality of liquid capsule grooves, and the orthographic projection of the liquid capsules on the first seat surface along the thickness direction of the substrate covers the liquid capsule grooves; the mixing chamber is arranged on the side of the first seat surface away from the second seat surface and is connected to the sample filling chamber through a microfluidic channel. The mixing chamber is connected with at least three liquid capsule grooves through microfluidic channels; the first pressurizing chamber is arranged on the side of the first seat surface away from the second seat surface and is connected with the mixing chamber through the microfluidic channel; the column mold cavity is arranged on the side of the first seat surface away from the second seat surface and is connected with the mixing chamber through the microfluidic channel, and a column mold is arranged in the column mold cavity; the amplification chamber is arranged on the side of the first seat surface away from the second seat surface and is connected with the column mold cavity through the microfluidic channel; the hybridization chamber is arranged on the side of the first seat surface away from the second seat surface, and the hybridization chamber is connected with the amplification chamber and part of the liquid capsule grooves through the microfluidic channel; wherein a drainage hole connecting the hybridization chamber and the second seat surface is arranged on the substrate; a coating is arranged on the side of the first seat surface away from the second seat surface, and the coating covers the mixing chamber, the amplification chamber and the hybridization chamber.
[0005] Optionally, the nucleic acid detection card box also includes multiple sets of valve structures, which are respectively arranged at the input and output ends of the amplification chamber, between the mixing chamber and the corresponding liquid capsule, and at the drainage hole to open or block the corresponding microchannel and the corresponding drainage hole.
[0006] Optionally, the nucleic acid detection card box also includes a plurality of convex cones, and the plurality of convex cones are arranged in a one-to-one correspondence in the plurality of liquid capsule grooves.
[0007] Optionally, the tip of the convex cone is perpendicular to the thickness of the substrate; the liquid capsule is a multi-layer structure and is made of metal foil.
[0008] Optionally, the nucleic acid detection card box also includes a plurality of freeze-dried reagent chambers, and the plurality of freeze-dried reagent chambers are respectively arranged on a microfluidic channel connecting the hybridization chamber and a liquid capsule tank.
[0009] Optionally, the nucleic acid detection card box also includes a heating component, which is arranged on the substrate and corresponds to the mixing chamber, the amplification chamber and the hybridization chamber, and the heating component heats the mixing chamber, the amplification chamber and the hybridization chamber in response to the driving force.
[0010] Optionally, the mixing chamber includes a first mixing chamber and a second mixing chamber, the first mixing chamber is connected to the sample filling chamber, the first pressurizing chamber and the liquid capsule groove corresponding to the liquid capsule filled with cleaning liquid and eluent through a microfluidic channel; the second mixing chamber is connected to the first mixing chamber, the column mold cavity and the liquid capsule groove corresponding to the liquid capsule filled with lysis liquid through a microfluidic channel.
[0011] Optionally, the nucleic acid detection card box also includes a second pressurizing chamber, which is arranged on the side of the first seat surface away from the second seat surface and connects the microchannel between the column mold cavity and the amplification chamber; wherein the connection area between the second pressurizing chamber and the corresponding microchannel is located between the valve structure at the input end of the amplification chamber and the amplification chamber.
[0012] Optionally, the nucleic acid detection card box also includes an exhaust flow channel, which connects the hybridization chamber and the first seat surface.
[0013] Optionally, the nucleic acid detection card box also includes a membrane chip, and the membrane chip is arranged on the inner wall of the hybridization chamber.
[0014] A nucleic acid detection cartridge is proposed in an embodiment of the present application, wherein the liquid capsules corresponding to the mixing chamber are respectively filled with a lysis solution, a cleaning solution and an elution solution, and the liquid capsules corresponding to the hybridization chamber are filled with reagents corresponding to each hybridization step, the nucleic acid sample is injected into the mixing chamber through the sample filling chamber, the lysis solution liquid capsule corresponding to the mixing chamber is squeezed, so that the lysis solution and the nucleic acid sample are mixed in the mixing chamber to form a first mixed solution, an air pump is connected to the first pressurizing chamber for pressurization, thereby pushing the first mixed solution through the column mold cavity and being filtered by the column mold, the nucleic acid and some impurities are retained by the column mold, the excess waste liquid enters the hybridization chamber after passing through the amplification chamber, and is discharged from the drainage hole at the hybridization chamber, then the cleaning liquid liquid capsule corresponding to the mixing chamber is squeezed, and the cleaning liquid is pushed through the column mold in the column mold cavity through the first pressurizing chamber to remove the impurities. The nucleic acid is washed down and discharged from the drainage hole, and then the eluent liquid capsule corresponding to the mixing chamber is squeezed, and the eluent is pushed through the column mold in the column mold cavity by the first pressurized chamber to elute the nucleic acid to form a second mixed liquid, and the nucleic acid extraction is completed; the first pressurized chamber pushes the second mixed liquid into the amplification chamber for amplification; after the amplification is completed, the drainage hole can be temporarily closed, and the liquid capsule corresponding to the hybridization chamber can be squeezed to allow the hybridization reagent to enter the hybridization chamber, and then the amplification product is pushed into the hybridization chamber for hybridization through the first pressurized chamber. Whatever reagent is needed during the hybridization process can be squeezed by squeezing the corresponding liquid capsule, and the waste liquid generated during the hybridization process can be discharged through the drainage hole, so that the extraction, amplification and hybridization operations of the nucleic acid can be completed, the operation is simple and efficient, no liquid transfer is required during the process, and it is difficult to cause pollution. BRIEF DESCRIPTION OF THE DRAWINGS
[0015] Figure 1 A schematic diagram of the overall structure of a nucleic acid detection cartridge provided in an embodiment of the present application;
[0016] Figure 2 for Figure 1 A schematic diagram of the structural disassembly of the embodiment in FIG.
[0017] Figure 3 for Figure 1 Another perspective structural schematic diagram of the embodiment in FIG.
[0018] In the figure: 1, substrate; 11, drainage hole; 12, liquid capsule groove; 121, convex cone; 13, valve structure; 2, sample filling chamber; 4, liquid capsule; 5, mixing chamber; 51, first mixing chamber; 52, second mixing chamber; 6, first pressurizing chamber; 7, column mold cavity; 8, amplification chamber; 81, second pressurizing chamber; 9, hybridization chamber; 91, freeze-dried reagent chamber; 92, exhaust flow channel.
[0019] The realization of the purpose, functional features and advantages of this application will be further explained in conjunction with embodiments and with reference to the accompanying drawings. DETAILED DESCRIPTION
[0020] The following will be combined with the drawings in the embodiments of the utility model to clearly and completely describe the technical solutions in the embodiments of the utility model. Obviously, the described embodiments are only part of the embodiments of the utility model, not all of the embodiments. Based on the embodiments of the utility model, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the utility model.
[0021] It should be noted that all directional indications in the embodiments of the present invention (such as up, down, left, right, front, back...) are only used to explain the relative position relationship, movement status, etc. between the components under a certain specific posture (as shown in the accompanying drawings). If the specific posture changes, the directional indication will also change accordingly.
[0022] In the present invention, unless otherwise clearly specified and limited, the terms "connection", "fixation", etc. should be understood in a broad sense. For example, "fixation" can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection or an indirect connection through an intermediate medium, it can be the internal connection of two elements or the interaction relationship between two elements, unless otherwise clearly defined. For ordinary technicians in this field, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.
[0023] In addition, if there are descriptions involving "first", "second", etc. in the embodiments of the utility model, the descriptions of "first", "second", etc. are only used for descriptive purposes and cannot be understood as indicating or implying their relative importance or implicitly indicating the number of technical features indicated. Therefore, the features defined as "first" and "second" may explicitly or implicitly include at least one of the features. In addition, the meaning of "and / or" appearing in the full text includes three parallel schemes. Taking "A and / or B" as an example, it includes scheme A, or scheme B, or a scheme that satisfies both A and B. In addition, the technical solutions between the various embodiments can be combined with each other, but it must be based on the ability of ordinary technicians in this field to implement. When the combination of technical solutions is contradictory or cannot be implemented, it should be deemed that such a combination of technical solutions does not exist and is not within the scope of protection required by the utility model.
[0024] refer to Figure 1 to Figure 3 It should be understood that Figure 2 The connections between the components should be like Figure 1 As compact as in the example, this is just an example. Figure 2Some components are disassembled and displayed for easy understanding. An embodiment of the present application provides a nucleic acid detection card box, which may include a substrate 1, a sample filling chamber 2, a plurality of liquid capsules 4, a mixing chamber 5, a first pressurized chamber 6, a column mold cavity 7, an amplification chamber 8 and a hybridization chamber 9. The substrate 1 has a first seat surface and a second seat surface that are arranged opposite to each other in the thickness direction thereof, and a plurality of liquid capsule grooves 12 are arranged on the side of the first seat surface away from the second seat surface; the sample filling chamber 2 is arranged on the side of the first seat surface away from the second seat surface; the plurality of liquid capsules 4 are all arranged on the side of the first seat surface away from the second seat surface and correspond one by one to the plurality of liquid capsule grooves 12, and the orthographic projection of the liquid capsule 4 on the first seat surface along the thickness direction of the substrate 1 covers the liquid capsule groove 12; the mixing chamber 5 is arranged on the side of the first seat surface away from the second seat surface and corresponds to the sample filling chamber 2. The mixing chamber 5 is connected with at least three liquid capsule grooves 12 through microfluidic channels; the first pressurizing chamber 6 is arranged on the side of the first seat surface away from the second seat surface and is connected with the mixing chamber 5 through the microfluidic channel; the column mold cavity 7 is arranged on the side of the first seat surface away from the second seat surface and is connected with the mixing chamber 5 through the microfluidic channel, and a column mold is arranged in the column mold cavity 7; the amplification chamber 8 is arranged on the side of the first seat surface away from the second seat surface and is connected with the column mold cavity 7 through the microfluidic channel; the hybridization chamber 9 is arranged on the side of the first seat surface away from the second seat surface, and the hybridization chamber 9 is connected with the amplification chamber 8 and part of the liquid capsule grooves 12 through the microfluidic channel; wherein, a drainage hole 11 connecting the hybridization chamber 9 and the second seat surface is arranged on the substrate 1; a coating is arranged on the side of the first seat surface away from the second seat surface, and the coating covers the mixing chamber 5, the amplification chamber 8 and the hybridization chamber 9.
[0025] A nucleic acid detection card box proposed in an embodiment of the present application has a liquid capsule 4 corresponding to the mixing chamber 5, which is respectively filled with a lysis solution, a cleaning solution and an elution solution, and a liquid capsule 4 corresponding to the hybridization chamber 9 is filled with reagents corresponding to each hybridization step. The nucleic acid sample is injected into the mixing chamber 5 through the sample filling chamber 2, and the lysis liquid capsule corresponding to the mixing chamber 5 is squeezed to mix the lysis liquid and the nucleic acid sample in the mixing chamber 5 to form a first mixed liquid. An air pump is connected to the first pressurizing chamber 6 for pressurization, thereby pushing the first mixed liquid through the column mold cavity 7 and being filtered by the column mold. The nucleic acid and some impurities are retained by the column mold, and the excess waste liquid enters the hybridization chamber 9 after passing through the amplification chamber 8 and is discharged from the drainage hole 11 at the hybridization chamber 9. Then, the cleaning liquid capsule corresponding to the mixing chamber 5 is squeezed, and the cleaning liquid is pushed through the column mold in the column mold cavity 7 through the first pressurizing chamber 6 to clean the impurities and discharge them from the drainage hole 11, and then the mixing is squeezed. The eluent capsule corresponding to the cavity 5 is pushed through the column mold in the column mold cavity 7 by the first pressurized chamber 6 to elute the nucleic acid to form a second mixed liquid, thereby completing the nucleic acid extraction; the first pressurized chamber 6 pushes the second mixed liquid into the amplification chamber 8 for amplification; after the amplification is completed, the drainage hole 11 can be temporarily closed, and the capsule 4 corresponding to the hybridization chamber 9 can be squeezed to allow the hybridization reagent to enter the hybridization chamber 9, and then the amplification product can be pushed into the hybridization chamber 9 for hybridization through the first pressurized chamber 6. What kind of reagent is needed during the hybridization process can be squeezed by squeezing the corresponding capsule 4, so that all the steps required for the hybridization reaction are gradually completed: hybridization, enzyme labeling, color development, cleaning, etc. At the same time, the waste liquid generated in each step is discharged through the drainage hole 11 at the bottom of the hybridization chamber 9, so that the extraction, amplification and hybridization operations of the nucleic acid can be completed, the operation is simple and efficient, no liquid transfer is required during the process, and it is difficult to cause pollution.
[0026] It should be noted that if Figure 1 As shown, the sample filling chamber 2, mixing chamber 5, first pressurizing chamber 6, column mold cavity 7, amplification chamber 8, hybridization chamber 9 and each microfluidic channel on the substrate 1 are arranged in the form of grooves on the side of the first side of the substrate 1 away from the second side. After the first side of the substrate 1 is covered with a coating, the mixing chamber 5, column mold cavity 7, amplification chamber 8, hybridization chamber 9 and each corresponding microfluidic channel located on the first side form a channel. At this time, the sample filling chamber 2 is connected to the outside world for filling samples, and can be closed after the filling is completed. The first pressurizing chamber 6 is connected to the outside world for connecting an air pump.
[0027] Among them, the microchannels connected to the column mold cavity 7 are all arranged on the side of the second seat surface away from the first seat surface. When in use, the thickness direction of the substrate 1 is the same as the gravity direction, and the first side surface is located above the second side surface. In this way, the first pressurizing chamber 6 applies gas pressure to the mixing chamber 5 to facilitate pushing the liquid in the mixing chamber 5 from the bottom of the mixing chamber 5 into the microchannel connected to the column mold cavity 7.
[0028] It should be understood that the lysing liquid capsule refers to the capsule 4 filled with lysing liquid, the cleaning liquid capsule refers to the capsule 4 filled with cleaning liquid, and the eluent liquid capsule refers to the capsule 4 filled with eluent.
[0029] Furthermore, the microfluidic chip, i.e., the nucleic acid detection card box in the present application, is made of polycarbonate or similar materials with certain transparency, toughness, strength, and high temperature resistance. The coating is also a transparent coating, which makes it convenient for staff to observe and take pictures. At the same time, the coating is coated with a material with good biocompatibility and can be directly pressed onto the first side of the substrate 1.
[0030] Among them, when the nucleic acid undergoes PCR (polymerase chain reaction) amplification reaction in the amplification chamber 8, since there is only one layer of film on the amplification chamber 8, the heating and cooling rate is increased compared to the microfluidic chip with a lid or a PCR tube, with an average rate of 5°C per second and a maximum rate of 10°C per second, thereby increasing the PCR amplification reaction rate.
[0031] Furthermore, when amplification is performed, the fourth pump membrane and the fifth pump membrane should both be in a closed state, so that the amplification chamber 8 is in a sealed state, thereby effectively preventing PCR amplification aerosol contamination.
[0032] refer to Figure 3 In an exemplary embodiment, the nucleic acid detection card box may also include multiple sets of valve structures 13, which are respectively arranged at the input end and the output end of the amplification chamber 8, between the mixing chamber 5 and the corresponding liquid capsule 4, and at the drainage hole 11 to open or block the corresponding microchannel and the corresponding drainage hole 11; the nucleic acid detection card box may also include an exhaust flow channel 92, which connects the hybridization chamber 9 and the first seat surface.
[0033] Furthermore, the valve structure 13 can be a common valve or a membrane pump. The membrane pump can drive the coating to fit into the microchannel side wall at the corresponding position or separate from the microchannel side wall at the corresponding position, so as to cut off or open the corresponding microchannel; the following description will be made by opening the membrane pump to open the corresponding microchannel and closing the membrane pump to cut off the corresponding microchannel, and the drain hole 11 will be opened and closed by a switch valve corresponding to the drain hole 11.
[0034] Among them, for the convenience of distinction, the membrane pump between the lysis liquid capsule and the mixing chamber 5 is recorded as the first membrane pump, the membrane pump between the cleaning liquid capsule and the mixing chamber 5 is recorded as the second membrane pump, the membrane pump between the elution liquid capsule and the mixing chamber 5 is recorded as the third membrane pump, the membrane pump between the column mold cavity 7 and the amplification cavity 8 is recorded as the fourth membrane pump, and the membrane pump between the amplification cavity 8 and the hybridization cavity 9 is recorded as the fifth membrane pump.
[0035] In the embodiment of the present application, before use, each membrane pump is in a closed state and the valve is in an open state. The nucleic acid sample is injected into the mixing chamber 5 through the sample filling chamber 2, the first membrane pump is turned on, and the lysis liquid capsule corresponding to the mixing chamber 5 is squeezed to mix the lysis liquid and the nucleic acid sample in the mixing chamber 5 to form a first mixed liquid. The fourth membrane pump and the fifth membrane pump are turned on, and the air pump is connected to the first pressurizing chamber 6 for pressurization, thereby pushing the first mixed liquid through the column mold cavity 7 and being filtered by the column mold. The nucleic acid and some impurities are retained by the column mold, and the excess waste liquid enters the hybridization chamber 9 after passing through the amplification chamber 8 and is discharged from the drainage hole 11 at the hybridization chamber 9. The second membrane pump is turned on, and then the cleaning liquid capsule corresponding to the mixing chamber 5 is squeezed, and the cleaning liquid is pushed through the column mold in the column mold cavity 7 through the first pressurizing chamber 6 to clean the impurities and The liquid is discharged through the drain hole 11, the third membrane pump is turned on, the valve is closed, and then the eluent capsule corresponding to the mixing chamber 5 is squeezed, and the eluent is pushed through the column mold in the column mold cavity 7 through the first pressurized chamber 6 to elute the nucleic acid to form a second mixed liquid, and the nucleic acid extraction is completed; the first pressurized chamber 6 pushes the second mixed liquid into the amplification chamber 8 for amplification; the liquid capsule 4 corresponding to the hybridization chamber 9 is squeezed to allow the hybridization reagent to enter the hybridization chamber 9, and then the amplification product is pushed into the hybridization chamber 9 for hybridization through the first pressurized chamber 6. Whatever reagent is needed during the hybridization process can be squeezed by squeezing the corresponding liquid capsule 4, and the waste liquid generated during the hybridization process can be discharged through the drain hole 11, so that the extraction, amplification and hybridization operations of nucleic acids can be completed, the operation is simple and efficient, no liquid transfer is required during the process, and it is difficult to cause pollution. Among them, the exhaust flow channel 92 is used to discharge excess air in each flow channel and chamber.
[0036] refer to Figure 2 In an exemplary embodiment, the nucleic acid detection cartridge may further include a plurality of convex cones 121 , and the plurality of convex cones 121 are disposed in a one-to-one correspondence within the plurality of liquid capsule grooves 12 .
[0037] Specifically, when the liquid capsule 4 is pressed, the side of the liquid capsule 4 facing the substrate 1 contacts the convex cone 121 , so that the liquid capsule 4 is punctured by the convex cone 121 , and the corresponding reagent in the liquid capsule 4 flows into the liquid capsule groove 12 .
[0038] Further, such as Figure 2 As shown, the tip of the convex cone 121 is oriented perpendicularly to the thickness of the substrate 1; the liquid capsule 4 is a multi-layer structure and is made of metal foil, which not only ensures that the liquid capsule 4 has a certain ductility, but also achieves the shape fixity of the liquid capsule 4 before and after squeezing, which is convenient for pre-setting various reagents. At the same time, when the staff squeezes the liquid capsule 4, the convex cone 121 is oriented perpendicularly to the thickness direction of the substrate 1, and the staff will not be injured.
[0039] refer to Figure 1 to Figure 3 In an exemplary embodiment, the nucleic acid detection card box may also include a plurality of freeze-dried reagent chambers 91 , and the plurality of freeze-dried reagent chambers 91 are respectively arranged on a microfluidic channel connecting the hybridization chamber 9 and a liquid capsule tank 12 .
[0040] like Figure 1 to Figure 3 As shown, there are two freeze-dried reagent chambers 91. After the reagent in the liquid capsule 4 connected to the freeze-dried reagent chamber 91 is squeezed out, the reagent liquid will be buffered in the corresponding freeze-dried reagent chamber 91. At the same time, the reagent will dissolve and flush the freeze-dried reagent, and then inject them into the hybridization chamber 9 together. In this way, the nucleic acid detection card box can be equipped with freeze-dried reagents and liquid reagents for hybridization at the same time, and the scope of use is wider.
[0041] In an exemplary embodiment, the nucleic acid detection card box may also include a heating component, which is arranged on the substrate 1 and corresponds to the mixing chamber 5, the amplification chamber 8 and the hybridization chamber 9, and the heating component heats the mixing chamber 5, the amplification chamber 8 and the hybridization chamber 9 in response to the driving force.
[0042] It should be noted that the heating component is not limited here, and a specific position on the substrate 1 can be heated to achieve temperature control. For example, the heating component can be an electric heating plate, which is attached to the second side away from the first side and corresponds one by one to the mixing chamber 5, the amplification chamber 8 and the hybridization chamber 9.
[0043] In the amplification chamber 8, the temperature cycle is controlled by an electric heating plate, and the cycle is performed 25 to 40 times in the order of 95°C, 55°C, and 68°C. The time length of each temperature section can be 5 seconds to 30 seconds; in the hybridization chamber 9, the 45°C required for hybridization, the 42°C required for enzyme labeling, and the 37°C required for color development can be controlled accordingly by the electric heating plate.
[0044] refer to Figure 1 to Figure 3 In an exemplary embodiment, the mixing chamber 5 may include a first mixing chamber 51 and a second mixing chamber 52. The first mixing chamber 51 is connected to the sample filling chamber 2, the first pressurizing chamber 6, and the liquid capsule groove 12 corresponding to the liquid capsule 4 filled with cleaning liquid and eluent through a microfluidic channel; the second mixing chamber 52 is connected to the first mixing chamber 51, the column mold cavity 7, and the liquid capsule groove 12 corresponding to the liquid capsule 4 filled with lysis liquid through a microfluidic channel.
[0045] It should be understood that if Figure 3 As shown, the second mixing chamber 52 is also connected to the third pressurizing chamber. After the lysate flows into the second mixing chamber 52, the fourth membrane pump is closed, and the pressurization through the third pressurizing chamber will cause the lysate to enter the first mixing chamber 51 and mix with the nucleic acid sample to form a first mixed liquid. Thereafter, the fourth membrane pump and the fifth membrane pump are turned on, and the first mixed liquid can pass through the column mold cavity 7 through the first pressurizing chamber 6 and the third pressurizing chamber.
[0046] refer to Figure 3In an exemplary embodiment, the nucleic acid detection card box may also include a second pressurizing chamber 81, which is arranged on the side of the first seat surface away from the second seat surface and connects the microchannel between the column mold cavity 7 and the amplification chamber 8; wherein the connection area between the second pressurizing chamber 81 and the corresponding microchannel is located between the valve structure 13 at the input end of the amplification chamber 8 and the amplification chamber 8.
[0047] like Figure 3 As shown, after the amplification is completed and before the PCR amplification product is pushed into the hybridization chamber 9, the fourth membrane pump and the valve are in a closed state, and the fifth membrane pump is in an open state. At this time, the PCR amplification product can be pushed into the hybridization chamber 9 through the second pressurized chamber 81. The second pressurized chamber 81 is closer to the PCR amplification product than the first pressurized chamber 6, and it is more convenient to push the PCR amplification product to move.
[0048] In an exemplary embodiment, the nucleic acid detection cartridge may further include a membrane chip, which is disposed on the inner wall of the hybridization chamber 9 .
[0049] Specifically, after the hybridization reaction is completed, the target DNA (deoxyribonucleic acid) will combine with the probe preset on the membrane chip to develop color, thereby detecting whether the sample contains the target DNA.
[0050] When conducting hybridization reaction, each reagent liquid should immerse the membrane chip. When the entire hybridization process is completed, the final result can be displayed on the membrane chip. The membrane chip can then be photographed, stored, identified, and interpreted, or the result can be observed and judged by human eyes.
[0051] The above are only preferred embodiments of the present application, and are not intended to limit the patent scope of the present application. Any equivalent structure or equivalent process transformation made using the contents of the present application specification and drawings, or directly or indirectly applied in other related technical fields, are also included in the patent protection scope of the present application.
Claims
1. A nucleic acid detection cartridge, characterized in that: include: A substrate (1) having a first seat surface and a second seat surface arranged opposite to each other in the thickness direction thereof, wherein a plurality of liquid capsule grooves (12) are arranged on a side of the first seat surface facing away from the second seat surface; A sample filling chamber (2) is arranged on a side of the first seat surface facing away from the second seat surface; A plurality of liquid capsules (4), each of which is arranged on a side of the first seat surface away from the second seat surface and corresponds one-to-one to the plurality of liquid capsule grooves (12), wherein the orthographic projection of the liquid capsule (4) on the first seat surface along the thickness direction of the substrate (1) covers the liquid capsule groove (12); A mixing chamber (5), arranged on a side of the first seat surface facing away from the second seat surface and connected to the sample filling chamber (2) through a microchannel, and the mixing chamber (5) is connected to at least three of the liquid capsule grooves (12) through the microchannel; A first pressurizing chamber (6), arranged on a side of the first seat surface away from the second seat surface and connected to the mixing chamber (5) through a microchannel; A column mold cavity (7) is arranged on a side of the first seat surface away from the second seat surface and is connected to the mixing cavity (5) through a microchannel, and a column mold is arranged in the column mold cavity (7); an amplification chamber (8), arranged on a side of the first seat surface facing away from the second seat surface and connected to the column mold cavity (7) through a microchannel; A hybridization chamber (9) is arranged on a side of the first seat surface facing away from the second seat surface, and the hybridization chamber (9) is connected to the amplification chamber (8) and part of the liquid capsule groove (12) through a microfluidic channel; Wherein, the substrate (1) is provided with a drainage hole (11) connecting the hybridization chamber (9) and the second seat surface; A coating is provided on the side of the first seat surface facing away from the second seat surface, and the coating covers the mixing chamber (5), the amplification chamber (8) and the hybridization chamber (9).
2. The nucleic acid detection cartridge according to claim 1, characterized in that: The nucleic acid detection cartridge further comprises: A plurality of valve structures (13) are respectively arranged at the input end and the output end of the amplification chamber (8), between the mixing chamber (5) and the corresponding liquid capsule (4), and at the drainage hole (11) to open or block the corresponding microchannel and the corresponding drainage hole (11).
3. The nucleic acid detection cartridge according to claim 1, characterized in that: The nucleic acid detection cartridge further comprises: A plurality of convex cones (121) are arranged in a one-to-one correspondence in the plurality of liquid capsule grooves (12).
4. The nucleic acid detection cartridge according to claim 3, characterized in that: The tip of the convex cone (121) is oriented in a direction perpendicular to the thickness of the substrate (1); The liquid capsule (4) is a multi-layer structure and is made of metal foil.
5. The nucleic acid detection cartridge according to claim 1, characterized in that: The nucleic acid detection cartridge further comprises: A plurality of freeze-dried reagent chambers (91) are respectively arranged on a microfluidic channel connecting the hybridization chamber (9) and one of the liquid capsule tanks (12).
6. The nucleic acid detection cartridge according to claim 1, characterized in that: The nucleic acid detection cartridge further comprises: A heating component is arranged on the substrate (1) and corresponds to the mixing chamber (5), the amplification chamber (8) and the hybridization chamber (9), and the heating component heats the mixing chamber (5), the amplification chamber (8) and the hybridization chamber (9) in response to a driving force.
7. The nucleic acid detection cartridge according to claim 1, characterized in that: The mixing chamber (5) comprises: The first mixing chamber (51) is connected to the sample filling chamber (2), the first pressurizing chamber (6) and the liquid capsule groove (12) corresponding to the liquid capsule (4) containing the cleaning liquid and the eluent through a microchannel; The second mixing chamber (52) is connected to the first mixing chamber (51), the column mold cavity (7) and the liquid capsule groove (12) corresponding to the liquid capsule (4) containing the lysis solution through a microchannel.
8. The nucleic acid detection cartridge according to claim 2, characterized in that: The nucleic acid detection cartridge further comprises: A second pressurizing chamber (81) is arranged on a side of the first seat surface away from the second seat surface and is connected to the microchannel between the column mold cavity (7) and the amplification cavity (8); Wherein, the connection area between the second pressurizing chamber (81) and the corresponding microfluidic channel is located between the valve structure (13) at the input end of the amplification chamber (8) and the amplification chamber (8).
9. The nucleic acid detection cartridge according to claim 1, wherein: The nucleic acid detection cartridge further comprises: An exhaust flow channel (92) connects the hybridization chamber (9) and the first seat surface.
10. The nucleic acid detection cartridge according to claim 1, characterized in that: The nucleic acid detection cartridge further comprises: The membrane chip is arranged on the inner wall of the hybridization chamber (9).