A microfluidic nucleic acid detection kit and a detection device

Abstract

The application discloses a micro-fluidic nucleic acid detection kit and a detection device. The first aspect provides a micro-fluidic nucleic acid detection kit, which comprises a reagent card and a box body capable of sliding relative to the reagent card to control the operation of the kit, and further comprises a gas source cavity, a liquid cavity, a nucleic acid amplification cavity and a gas tail cavity capable of controlling the gas pressure of a closed system through volume change. The gas tail cavity is communicated with the nucleic acid amplification cavity through a waste gas flow channel, and the waste gas flow channel is provided with a overflow valve. The second aspect provides a detection device, which comprises the above-mentioned micro-fluidic nucleic acid detection kit, a first shell, a power module and a display module. The nucleic acid detection kit is detachably inserted into the first shell, and further comprises a first heating module and a first detection module. The third aspect provides an integrated detection device, which comprises the above-mentioned micro-fluidic nucleic acid detection kit and a second shell, a second heating module and a second detection module. The device can completely separate from laboratory detection, is fully closed and pollution-free, and does not use external power.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of in vitro diagnostic devices, in particular to a microfluidic nucleic acid detection kit and detection device that can be used in on-site instant detection scenarios. BACKGROUND

[0002] In vitro diagnostic (IVD) technology refers to obtaining relevant clinical diagnostic information by detecting samples such as blood, body fluid, and tissue outside the human body, thereby helping to judge the disease or function of the body. The IVD field involves a very wide range of products and services, including blood testing, clinical biochemistry, cell diagnosis, microbiological diagnosis, coagulation diagnosis, immunochemistry, molecular diagnosis, point-of-care testing (POCT), etc. At present, the most promising and fastest-growing IVD is molecular diagnosis and instant detection POCT technology, which mainly detects nucleic acids.

[0003] With the progress of society, there is an increasing demand for instant detection POCT diagnostic devices that can be used in scenarios outside the laboratory, are low-cost, and are safe, such as in community hospitals, pharmacies, and homes. At present, most nucleic acid detection kits on the market cannot be used outside professional biological laboratories. This is mainly because sample operation needs to be protected from contamination, so it must be completed in a laboratory, and the entire process requires several professional devices to complete the lysis of biological samples, the extraction of nucleic acid molecules, nucleic acid PCR amplification, and the collection of optical signals and data analysis.

[0004] In recent years, with the progress of science and technology, advanced nucleic acid detection integrated, fully automated POCT reagent kits and supporting equipment have gradually appeared in the industry, such as Xpert products of Cypheid, filmarray products of Biofire, and Vivalytic products of Bosch. These products use external power to operate the components in the reagent kit, realize the control of various liquid flows in the reagent kit, and realize the automation of the whole process of nucleic acid sample lysis, extraction, amplification and detection. But the detection instrument and reagent card using this way are expensive, and the equipment is still complex and heavy, not convenient to carry, most of which still need to be operated in a special PCR laboratory, and cannot truly realize the application scene of on-site instant detection. Although some POCT reagent kits use constant temperature amplification technology and one-step nucleic acid lysis and extraction technology to greatly simplify the reagent card and detection instrument, and even realize household nucleic acid detectors, such as the household nucleic acid detection product of Lucira company in the United States, which realizes sample extraction and amplification in the box without relying on external equipment, in order to realize accurate liquid flow and control, it is necessary to leave an air outlet on the reagent kit to realize liquid control. The filter membrane is selected and set on the air outlet to prevent aerosol pollution, but these filter membranes cannot completely prevent the overflow of aerosol, especially the LAMP constant temperature amplification technology used will cause the virus concentration in the cavity and gas after nucleic acid amplification to be very high. SUMMARY

[0005] The purpose of the present application is to provide an integrated, fully automated POCT microfluidic nucleic acid detection reagent kit and its detection device, to solve the technical problems that the reagent kit cannot be completely separated from the laboratory for detection, cannot be fully closed and pollution-free, and the device system is complex due to the use of external power.

[0006] As conceived above, the technical solution adopted by the first aspect of the present application is:

[0007] A microfluidic nucleic acid detection reagent kit, comprising a box body and a reagent card connected to each other, the box body being capable of sliding relative to the reagent card to control the operation of the reagent kit, further comprising a plurality of cavities and a plurality of flow channels for connecting the plurality of cavities, the plurality of cavities and the plurality of flow channels forming a closed system, wherein the plurality of cavities at least comprises:

[0008] A gas source cavity is arranged in the box body and can control the gas pressure of the closed system by changing the volume thereof;

[0009] A liquid cavity is arranged in the box body for injecting liquid reagent, and the liquid cavity and the gas source cavity are connected by a gas source flow channel;

[0010] A nucleic acid amplification cavity is arranged on the reagent card, and the nucleic acid amplification cavity is communicated with the liquid cavity through a sample inlet flow channel; a gas tail cavity is arranged on the reagent card, and the gas tail cavity is communicated with the nucleic acid amplification cavity through a waste gas flow channel; and a first sealing film is arranged on the reagent card, and the first sealing film seals the sample inlet flow channel, the waste gas flow channel and the gas tail cavity.

[0011] Further, a spill-proof valve is arranged at the waste gas flow channel, and the spill-proof valve is used for blocking the flow of liquid while being capable of discharging gas.

[0012] Further, the plurality of cavities further comprise a bubble storage cavity arranged between the waste gas flow channel and the nucleic acid amplification cavity, and the bubble storage cavity is communicated with the nucleic acid amplification cavity.

[0013] Further, the cartridge and the reagent card are connected through a buckle assembly, which comprises a buckle piece arranged on the reagent card and a buckle socket arranged on the cartridge.

[0014] The buckle piece comprises a buckle strip, one end of the buckle strip is connected with the reagent card, the other end of the buckle strip is a free end, and the buckle strip extends towards the cartridge, the buckle strip has a first cantilever, the reagent card has a second cantilever, the second cantilever is located above the first cantilever, and the buckle socket comprises a first buckle socket corresponding to the first cantilever and a second buckle socket corresponding to the second cantilever.

[0015] Further, before the cartridge slides relative to the reagent card, the first cantilever and the second cantilever are both in abutment with the cartridge, the gas pressure of the gas source cavity is the same as the gas pressure of the gas tail cavity, and the liquid in the liquid cavity does not flow into the nucleic acid amplification cavity.

[0016] When the cartridge slides relative to the reagent card, the gas source cavity is extruded, the gas pressure in the gas source cavity is increased, thereby pushing the liquid reagent in the liquid cavity to flow into the nucleic acid amplification cavity through the sample inlet flow channel, the flow of the liquid reagent pushes the gas in the nucleic acid amplification cavity to flow into the gas tail cavity, and when the liquid reagent in the liquid cavity reaches the spill-proof valve, the flow of the liquid reagent is stopped, thereby accurately injecting the liquid reagent into the nucleic acid amplification cavity.

[0017] Further, a solid reagent required for nucleic acid amplification is placed in the nucleic acid amplification cavity, the solid reagent comprises a dry powder reagent or a reagent freeze-dried ball or a quality control internal standard reagent; a nucleic acid lysis extraction liquid is placed in the liquid cavity, and a space required for adding a sample is reserved, and the volume of the nucleic acid lysis extraction liquid is 100 μL to 5000 μL.

[0018] Further, a main flow channel is arranged between the nucleic acid amplification cavity and the sample injection flow channel, and a wax column is arranged between the sample injection flow channel and the main flow channel; when the cartridge slides relative to the reagent card, the wax column is melted by a heating device to make the sample injection flow channel and the main flow channel communicate.

[0019] Further, the plurality of cavities further comprise a quality control internal standard cavity arranged in the cartridge, and a first lyophilized ball is arranged in the quality control internal standard cavity, and the first lyophilized ball contains reagent components required for internal quality control.

[0020] Further, a second lyophilized ball or lyophilized powder is arranged in the nucleic acid amplification cavity, and the second lyophilized ball or lyophilized powder contains reagent components required for nucleic acid amplification.

[0021] Further, the number of the nucleic acid amplification cavities, the overflow stop valves and the gas tail cavities are the same and correspond one by one.

[0022] Further, the bottom of the quality control internal standard cavity is inclined at an angle with respect to the horizontal plane, and the first lyophilized ball outlet is located at the lowest position of the bottom of the quality control internal standard cavity.

[0023] Further, one end of the sample injection flow channel close to the liquid cavity is provided with a puncture needle, and the puncture needle can be inserted into the liquid cavity to make the sample injection flow channel communicate with the liquid cavity.

[0024] Further, the reagent card is provided with a piston column, one end of the piston column is provided with a rubber piston, and the piston column is arranged in the cartridge.

[0025] The gas source cavity is in the form of a blind hole on the cartridge, one end of the piston column with the rubber piston extends into the blind end of the gas source cavity through the opening end of the gas source cavity, and the gas source flow channel is arranged close to the blind end of the gas source cavity.

[0026] Before the cartridge slides relative to the reagent card, the rubber piston is lower than the gas source flow channel in the gas source cavity.

[0027] When the cartridge slides relative to the reagent card, the rubber piston moves towards the blind end to compress and extrude the gas in the gas source cavity into the liquid cavity.

[0028] Further, a sample adding port is arranged on the cartridge, the sample adding port can communicate with the liquid cavity, one end of the liquid cavity away from the sample injection flow channel is provided with a sample adding port, the quality control internal standard cavity is provided with a first lyophilized ball inlet and a first lyophilized ball outlet, and the first lyophilized ball outlet communicates the liquid cavity and the quality control internal standard cavity.

[0029] Further, the box body further comprises an upper cover, the upper cover has a protrusion capable of sealing the sample adding port and the first freeze-dried ball outlet, and the upper cover is capable of covering the first freeze-dried ball inlet.

[0030] Further, one end of the liquid cavity away from the sample adding port is provided with a second sealing film, the liquid cavity is capable of being inserted with the puncture needle to make one end of the puncture needle pass through the second sealing film and communicate with the liquid cavity; the first freeze-dried ball inlet is provided with a third sealing film, and the first freeze-dried ball outlet is provided with a fourth sealing film; wherein,

[0031] The fourth sealing film comprises a first part, a second part and a third part which are integrally formed, the first part seals the first freeze-dried ball outlet, the second part seals the liquid cavity, and the third part is a pull ring structure and capable of extending out of the sample adding port.

[0032] Further, the diameter of the sample adding flow channel gradually tapers from one end away from the main flow channel to one end close to the main flow channel.

[0033] The nucleic acid detection kit provided in the first aspect of the application is completely closed and free of pollution, and is completely separated from laboratory use, because the whole process of sliding of the box body relative to the reagent card is carried out in the detection kit, and no gas outlet is formed on the detection kit; at the same time, the detection kit does not need external power, and the detection device is simple; the complete closure is conceived by the inventors based on the principle that gas can be compressed but liquid cannot be compressed: before the box body slides relative to the reagent card, the gas pressure of the gas source cavity and the gas pressure of the gas tail cavity are the same, and the second sealing film is not punctured, and the liquid in the liquid cavity will not flow into the nucleic acid amplification cavity; when the box body slides relative to the reagent card, the gas source cavity is extruded, the gas pressure in the gas source cavity is increased, thereby pushing the liquid reagent in the liquid cavity to inject into the nucleic acid amplification cavity through the sample adding flow channel, and the flow of the liquid reagent pushes the gas in the nucleic acid amplification cavity to inject into the gas tail cavity; when the liquid reagent in the liquid cavity reaches the overflow stop valve, the liquid reagent stops flowing, thereby accurately injecting the liquid reagent into the nucleic acid amplification cavity.

[0034] According to the second aspect of the application, the technical scheme adopted is:

[0035] A detection device is provided, comprising the microfluidic nucleic acid detection kit as described above, further comprising a first shell, a power module and a display module, the nucleic acid detection kit is detachably inserted on the first shell, and the detection device further comprises:

[0036] A first heating module is used to heat the nucleic acid amplification cavity.

[0037] a first detection module disposed in the first housing and configured to detect a change in a light signal in the nucleic acid amplification cavity, the light signal including one or more of a color, a brightness, a fluorescence, or a saturation signal.

[0038] Further, the nucleic acid amplification device further comprises a first circuit board disposed in the first housing and a control unit disposed on the first circuit board, the control unit being electrically connected to the first heating module to control the first heating module to heat the nucleic acid amplification cavity.

[0039] Further, the first heating module comprises a heating block closely surrounding the nucleic acid amplification cavity, a first heating sheet disposed at a bottom of the heating block and in contact with the heating block to conduct heat, and a temperature sensor adjacent to the first heating sheet, the temperature sensor being electrically connected to the control unit, and a heat dissipation space being formed between the first heating module and the first circuit board.

[0040] Further, the heating block is provided with a heating groove, and a first hole and a second hole oppositely disposed, the first hole and the second hole both penetrating the heating groove, the nucleic acid amplification cavity being capable of extending into the heating groove and being located between the first hole and the second hole.

[0041] Further, the first detection module comprises a light source and a light sensor electrically connected to the first circuit board, the light source and the light sensor both being disposed outside the heating block, the light source being in contact with the first hole, and the light sensor being in contact with the second hole.

[0042] Further, the first circuit board is provided with a processing unit, the processing unit being configured to process and analyze the light signal collected by the detection module, and then feed back the analysis result to the display module.

[0043] Further, the processing unit comprises a data acquisition circuit, an ADC chip, and a processor, the data acquisition circuit being configured to acquire a signal of the light sensor, the signal being converted into a digital signal by the ADC chip, and then being processed and analyzed by the processor, and then the analysis result being fed back to the display module to obtain detection data.

[0044] Further, the first circuit board is provided with a data communication unit, the data communication unit being configured to transmit the detection data to a wireless terminal, the wireless terminal including one or more of a mobile phone terminal, a computer, or a network server.

[0045] Further, the first shell comprises a shell body with an opening at the upper end and a shell upper cover rotatably connected with the shell body and used for covering the opening, the nucleic acid detection kit is detachably plugged at the opening, and the shell body is provided with a mounting position for mounting the display module, and the display module is arranged on the mounting position.

[0046] The second aspect of the present application provides a detection device, wherein the nucleic acid detection kit is detachably plugged on the detection device, and the detection kit is replaced after one-time use,

[0047] The nucleic acid detection device is a nucleic acid detection device completely separated from the laboratory, and full-closed sample detection is achieved, and the pollution risk of aerosol is completely eliminated.

[0048] According to the third aspect of the present application, the technical scheme adopted is as follows:

[0049] An integrated detection device is provided, comprising the microfluidic nucleic acid detection kit of the first aspect and a second shell, and further comprising:

[0050] A second heating module is used for heating the nucleic acid amplification cavity;

[0051] A second detection module is arranged in the second shell and used for detecting the change of the optical signal in the nucleic acid amplification cavity, and the optical signal comprises one or more signals of color, brightness, fluorescence or saturation.

[0052] Further, the shell can be integrally formed or the first shell comprises a front cover and a rear cover which are detachably connected, and the front cover and the rear cover are used for cooperating to clamp the microfluidic nucleic acid detection kit, and the second detection module is arranged between the front cover and the rear cover.

[0053] Further, a second circuit board arranged between the front cover and the rear cover and a control unit arranged on the second circuit board are further included, the control unit is electrically connected with the second heating module to control the second heating module to heat the nucleic acid amplification cavity, and a power socket is further arranged on the second circuit board.

[0054] Further, the second heating module comprises a second heating sheet closely surrounding the nucleic acid amplification cavity and a temperature sensor arranged on the inner surface of the second heating sheet and in contact with the reagent card, and the temperature sensor is electrically connected with the control unit.

[0055] Further, the second detection module comprises a light source and a light guide column which are electrically connected with the second circuit board, and the light source and the light guide column are respectively located on two sides of the reagent card.

[0056] Further, the second circuit board has a processing unit, which is used for processing and analyzing the optical signal collected by the second detection module to obtain detection data.

[0057] Further, the processing unit comprises a data acquisition circuit, an ADC chip and a processor, the data acquisition circuit acquires the signal of the optical sensor, the signal is converted into digital signal by the ADC chip, and then the digital signal is processed and analyzed by the processor to obtain the detection data.

[0058] Further, the second circuit board has a data communication unit, which is used for transmitting the detection data to a mobile phone terminal, a computer, a network server or other wireless terminals.

[0059] The present application has the following advantages:

[0060] The nucleic acid detection kit provided by the present application is integrated with the detection device, so that the kit and the detection device are no longer separated, and an integrated and disposable nucleic acid detection kit is formed, which is an indivisible whole, and one person uses one kit, and the detection device is abandoned, so that the contamination of the detection device on the reagent card is prevented; meanwhile, the integrated nucleic acid detection device is a nucleic acid detection device completely separated from a laboratory, and the whole closed sampling detection is achieved, so that the pollution risk of aerosol is completely eliminated. BRIEF DESCRIPTION OF DRAWINGS

[0061] Figure 1 is a structural schematic view of the nucleic acid detection kit provided by the embodiment one of the present application;

[0062] Figure 2 is a sectional structural schematic view of the nucleic acid detection kit provided by the embodiment one of the present application (the box body is not slid relative to the reagent card);

[0063] Figure 3 is another sectional structural schematic view of the nucleic acid detection kit provided by the embodiment one of the present application (the box body is not slid relative to the reagent card);

[0064] Figure 4 is a sectional structural schematic view of the nucleic acid detection kit provided by the embodiment one of the present application (the box body is slid relative to the reagent card); Figure 3 is a partial enlarged view of the B area in FIG. 8;

[0065] Figure 5 is a sectional structural schematic view of the nucleic acid detection kit provided by the embodiment one of the present application (the box body is slid relative to the reagent card);

[0066] Figure 6 is a sectional structural schematic view of the nucleic acid detection kit provided by the embodiment one of the present application (the box body is slid relative to the reagent card); Figure 5 is a partial enlarged view of the A area in FIG. 9;

[0067] Figure 7It is the perspective view of nucleic acid detection kit provided by the embodiment one of the present application;

[0068] Figure 8 It is the exploded view of nucleic acid detection kit provided by the embodiment one of the present application;

[0069] Figure 9 It is the air pressure balance method illustration of nucleic acid detection kit provided by the embodiment one of the present application;

[0070] Figure 10 It is the perspective view of nucleic acid detection device provided by the second embodiment of the present application;

[0071] Figure 11 It is the rear view structure schematic diagram of nucleic acid detection device provided by the second embodiment of the present application;

[0072] Figure 12 It is the cross-sectional view of nucleic acid detection device provided by the second embodiment of the present application;

[0073] Figure 13 It is the exploded view of nucleic acid detection device provided by the second embodiment of the present application;

[0074] Figure 14 It is the structure schematic diagram of heating block provided by the second embodiment of the present application;

[0075] Figure 15 It is the perspective view of integrated detection device provided by the third embodiment of the present application;

[0076] Figure 16 It is the exploded view of integrated detection device provided by the third embodiment of the present application;

[0077] Figure 17 It is the perspective view of front cover of integrated detection device provided by the third embodiment of the present application;

[0078] Figure 18 It is the perspective view of rear cover of integrated detection device provided by the third embodiment of the present application;

[0079] Figure 19 It is the structure schematic diagram of heating module of integrated detection device provided by the third embodiment of the present application.

[0080] In the figure:

[0081] 10, first housing; 11, housing body; 12, mounting base plate; 121, circuit board mounting seat; 122, back cover mounting base slot; 123, circuit board cover plate; 13, housing upper cover; 14, back cover; 15, debugging port; 16, first power socket; 17, power switch; 18, first LED lamp;

[0082] 21, nucleic acid detection kit; 22, first circuit board; 23, piston column; 24, rubber piston; 25, puncture needle; 26, second sealing film; 27, O-shaped sealing ring;

[0083] 211, reagent card; 2110, first sealing film; 2111, nucleic acid amplification cavity; 2112, gas tail cavity; 2113, sample inlet flow channel; 2114, main flow channel; 2115, waste gas flow channel; 2116, first branch flow channel; 2117, second branch flow channel; 2118, third branch flow channel; 2119, wax column; 2120, bubble storage cavity;

[0084] 212, box body; 2121, liquid cavity; 21211, first lyophilized ball inlet; 21212, first lyophilized ball outlet; 2122, gas source cavity; 2123, upper cover; 2124, sample addition port; 2125, protrusion; 2126, quality control internal standard cavity; 2127, third sealing film; 2128, fourth sealing film; 21281, first part; 21282, second part; 21283, third part; 2129, gas source flow channel;

[0085] 213, buckle assembly; 2131, buckle strip; 21311, first cantilever; 2132, bayonet buckle; 2133, second cantilever;

[0086] 3, first heating module; 31, heating block; 311, heating groove; 312, first hole; 32, first heating sheet;

[0087] 41, light source; 42, light sensor;

[0088] 51, front cover; 511, light guide column hole; 512, fixing column; 513, buckle assembly slot; 514, first bottom surface; 515, first side surface I; 516, second side surface I; 517, third side surface I; 52, rear cover; 521, fixing slot; 522, second bottom surface; 523, first side surface II; 524, second side surface II; 525, third side surface II;

[0089] 6, second heating module; 61, second heating sheet; 62, second circuit board; 621, second power socket; 63, temperature sensor; 64, third hole;

[0090] 71, light guide column; 72, second LED lamp;

[0091] 80, first lyophilized ball; 81, lyophilized ball one; 82, lyophilized ball two; 83, lyophilized ball three; 84, plastic ball;

[0092] 90, overflow valve; 91, overflow valve one; 92, overflow valve two; 93, overflow valve three. DETAILED DESCRIPTION

[0093] Embodiments of the present application are described in detail below with reference to the attached drawing figures, wherein the same or like reference numerals are used throughout the drawings to refer to the same or like elements or elements having the same or similar functionality. The following detailed description is exemplary in nature and is intended to provide a thorough and complete disclosure of the application, including its best mode(s), and is not intended to be limiting as to the scope or applicability of the application.

[0094] In the description of the present application, unless otherwise clearly specified and limited, the terms "connected", "connected", "fixed" should be understood broadly, for example, it can be fixed connection, or detachable connection, or integrated; it can be mechanical connection, or electrical connection; it can be directly connected, or indirectly connected through an intermediate medium, or 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 present application can be understood according to the specific circumstances.

[0095] In the present application, unless otherwise clearly specified and limited, the first feature "on" or "under" the second feature can include that the first and second features are in direct contact, or that the first and second features are not in direct contact but are in contact through another feature between them. Moreover, the first feature "on", "above" and "above" the second feature includes that the first feature is directly above and obliquely above the second feature, or only indicates that the horizontal height of the first feature is higher than that of the second feature. The first feature "below", "below" and "below" the second feature includes that the first feature is directly below and obliquely below the second feature, or only indicates that the horizontal height of the first feature is less than that of the second feature.

[0096] The technical solutions of the present application are further illustrated below in conjunction with the drawings and through specific embodiments.

[0097] Example one,

[0098] The nucleic acid detection kit provided by the embodiment of the present application is a nucleic acid detection kit 21 that can completely separate from the laboratory. Nucleic acid extraction is directly completed in the nucleic acid detection kit 21, and an internal air pressure balancing system is used in the nucleic acid detection kit 21 to store waste gas in the nucleic acid detection kit 21, so as to achieve completely closed sampling detection and completely eliminate the pollution risk of aerosol and the like. In addition, the sampling is pushed in the nucleic acid detection kit 21, which reduces the external power of the equipment and simplifies the complexity of the detection equipment.

[0099] ReferenceFigures 1-8 The microfluidic nucleic acid detection kit 21 comprises a box body 212 and a reagent card 211 connected with each other, the box body 212 can slide relative to the reagent card 211 to control the operation of the kit, and further comprises a plurality of cavities and a plurality of flow channels for connecting the plurality of cavities, the plurality of cavities and the plurality of flow channels form a closed system, wherein the plurality of cavities at least comprises:

[0100] A gas source cavity 2122 is arranged in the box body 212 and can control the gas pressure of the closed system by changing the volume thereof;

[0101] A liquid cavity 2121 is arranged in the box body 212 and used for injecting liquid reagent, the liquid cavity 2121 and the gas source cavity 2122 are connected through a gas source flow channel 2129;

[0102] A nucleic acid amplification cavity 2111 is arranged on the reagent card 211, the nucleic acid amplification cavity 2111 and the liquid cavity 2121 are connected through a sample injection flow channel 2113, and the internal gas pressure of the nucleic acid amplification cavity can be 0-100 kPa;

[0103] A gas tail cavity 2112 is arranged on the reagent card 211, the gas tail cavity 2112 and the nucleic acid amplification cavity 2111 are connected through a waste gas flow channel 2115, and the internal gas pressure of the gas tail cavity can be 0-100 kPa;

[0104] A bubble storage cavity 2120 is arranged between the waste gas flow channel 2115 and the nucleic acid amplification cavity 2111, the bubble storage cavity 2120 is connected with the nucleic acid amplification cavity 2111, and a spill-proof valve 90 is arranged between the bubble storage cavity 2120 and the waste gas flow channel 2115, the spill-proof valve 90 is suitable for blocking the flow of liquid and discharging gas.

[0105] Specifically, the plurality of cavities and the plurality of flow channels form the principle of the internal gas pressure balance process of the microfluidic nucleic acid detection kit 21, when the box body 212 slides relative to the reagent card 211, the gas pressure of the gas source cavity 2122 is the same as the gas pressure of the gas tail cavity 2112, and the liquid in the liquid cavity 2121 does not flow into the nucleic acid amplification cavity 2111;

[0106] When the box body 212 slides relative to the reagent card 211, the gas source cavity 2122 is extruded, the gas pressure in the gas source cavity 2122 is increased, thereby pushing the liquid reagent in the liquid cavity 2121 to flow into the nucleic acid amplification cavity 2111 through the sample injection flow channel 2113, and the flow of the liquid reagent pushes the gas in the nucleic acid amplification cavity 2111 to flow into the gas tail cavity 2112;

[0107] When the liquid reagent in the liquid cavity 2121 reaches the spill-proof valve 90, the flow of the liquid reagent is stopped, thereby accurately injecting the liquid reagent into the nucleic acid amplification cavity 2111;

[0108] Since the entire process of the box 212 sliding relative to the reagent card 211 takes place inside the nucleic acid test kit 21, there is no need to open an air vent on the nucleic acid test kit 21, which can achieve a completely sealed and pollution-free environment.

[0109] More specifically, the nucleic acid amplification chamber 2111 contains solid reagents required for nucleic acid amplification, including dry powder reagents, lyophilized reagent bulbs, or quality control internal standard reagents; the liquid chamber 2121 contains nucleic acid lysis extract and provides space for adding samples, wherein the volume of the liquid chamber 2121 is 100 μL to 3000 μL, and the volume of the nucleic acid lysis extract is 100 μL to 5000 μL.

[0110] To better illustrate the principle of the above liquid flow process, the following formula and process are used for explanation: Figure 2 , Figure 3 , Figure 5 and Figure 10 As shown: the volume of the gas source chamber 2122 is V1, the air volume of the liquid chamber 2121 is V2, the volume of the nucleic acid amplification chamber 2111 is V3, the volume of the gas tail chamber 2112 is V4, and the liquid flow formula is as follows:

[0111] Initial flow state of the lysis buffer:

[0112] The lysis fluid flow state begins with the push state:

[0113] Filtration fluid flow state: (The text appears to be incomplete and contains several grammatical errors. A more accurate translation would require the full context.)

[0114] During the sliding process of the box 212 relative to the reagent card 211, P1 is greater than P2, the flow state of the lysis solution changes to push liquid, P2 gradually increases until the liquid reagent in the liquid chamber 2121 reaches the overflow valve; the flow state of the lysis solution changes to stop liquid; at this time, P1 is still greater than P2.

[0115] like Figure 9 As shown, the following explanation uses the following case: the volume of the gas source chamber 2122 is 300 μl, the air volume of the liquid chamber 2121 is 800 μl, the volume of the nucleic acid amplification chamber 2111 is 80 μl, and the volume of the gas tail chamber 2112 is 500 μl.

[0116] Initial flow state of the lysis buffer:

[0117] The lysis fluid flow state begins with the push state:

[0118] The pyrolysis fluid flow state begins with the liquid stoppage state:

[0119] The main flow channel 2114 is arranged between the nucleic acid amplification cavity 2111 and the sample inlet flow channel 2113. Specifically, in order to enable the liquid cavity 2121 and the nucleic acid amplification cavity 2111 to be in a closed state during sample lysis, a wax column 2119 is arranged between the sample inlet flow channel 2113 and the main flow channel 2114 as a valve. When the box body 212 slides relative to the reagent card 211, the wax column 2119 is in a solid state, and the sample inlet flow channel 2113 and the main flow channel 2114 are blocked by the wax column 2119. The wax column 2119 is melted by a heating device, and the wax column 2119 changes phase from a solid state to a liquid state, so that the sample inlet flow channel 2113 and the main flow channel 2114 are communicated. The plurality of cavities further include a quality control internal standard cavity 2126 arranged in the box body 212. The quality control internal standard cavity 2126 contains a first freeze-dried ball 80. The first freeze-dried ball 80 contains reagent components required for internal quality control. The nucleic acid amplification cavity 2111 contains a second freeze-dried ball or freeze-dried powder. The second freeze-dried ball or freeze-dried powder contains reagent components required for nucleic acid amplification.

[0120] The nucleic acid amplification cavity 2111 and the gas tail cavity 2112 are the same in number and one-to-one correspondence. Similarly, a corresponding overflow stop valve 90 is arranged between each nucleic acid amplification cavity 2111 and the gas tail cavity 2112. Specifically, the number of the nucleic acid amplification cavity 2111 can be changed according to actual needs. For the convenience of description, in this embodiment, the nucleic acid amplification cavity 2111 includes a nucleic acid amplification cavity one, a nucleic acid amplification cavity two, and a nucleic acid amplification cavity three. Therefore, the corresponding gas tail cavity 2112 includes a gas tail cavity one, a gas tail cavity two, and a gas tail cavity three. At the same time, the corresponding overflow stop valve 90 includes an overflow stop valve one 91, an overflow stop valve two 92, and an overflow stop valve three 93. The overflow stop valve one 91 and the nucleic acid amplification cavity one form a gas bubble storage cavity one. The overflow stop valve two 92 and the nucleic acid amplification cavity two form a gas bubble storage cavity two. The overflow stop valve three 93 and the nucleic acid amplification cavity three form a gas bubble storage cavity three. The gas tail cavity 2112 and the nucleic acid amplification cavity 2111 are communicated through a waste gas flow channel 2115. That is, the overflow stop valve 90 is arranged on the waste gas flow channel 2115. Furthermore, the nucleic acid amplification cavity one and the main flow channel 2114 are communicated through a branch flow channel one 2116. The nucleic acid amplification cavity two and the main flow channel 2114 are communicated through a branch flow channel two 2117. The nucleic acid amplification cavity three and the main flow channel 2114 are communicated through a branch flow channel three 2118. The nucleic acid amplification cavity one pre-encapsulates a freeze-dried ball one 81. The nucleic acid amplification cavity two pre-encapsulates a freeze-dried ball two 82. The nucleic acid amplification cavity three pre-encapsulates a freeze-dried ball three 83. The freeze-dried ball one 81, the freeze-dried ball two 82, and the freeze-dried ball three 83 are all reagent systems (enzyme, template concentration, primer) required for amplification. The primers of the freeze-dried ball one 81, the freeze-dried ball two 82, and the freeze-dried ball three 83 are different, which are target point one primer of the target sample, target point two primer of the target sample, and primer of the internal standard, respectively. The internal pressure of the freeze-dried ball one 81, the freeze-dried ball two 82, and the freeze-dried ball three 83 can be 0-100 kPa.

[0121] The end of the sample flow channel 2113 close to the liquid cavity 2121 has a puncture needle 25, and the combination of the sample flow channel 2113 and the puncture needle 25 is a puncture tube. An O-shaped sealing ring 27 is arranged between the puncture tube and the extraction cavity. The puncture needle 25 can be inserted into the liquid cavity 2121, so that the sample flow channel 2113 communicates with the liquid cavity 2121. The diameter of the sample flow channel 2113 gradually decreases from the end away from the main flow channel 2114 to the end close to the main flow channel 2114. The reagent card 211 has a piston column 23, and one end of the piston column 23 is provided with a rubber piston 24.

[0122] The gas source cavity 2122 is a blind hole on the box body 212. The end of the piston column 23 with the rubber piston 24 extends into the blind end of the gas source cavity 2122 through the open end of the gas source cavity 2122. The gas source flow channel 2129 is arranged close to the blind end of the gas source cavity 2122. Before the box body 212 slides relative to the reagent card 211, the rubber piston 24 is lower than the gas source flow channel 2129 in the gas source cavity 2122. In this embodiment, the distance between the rubber piston 24 and the blind end is 1-10 mm, and preferably 5 mm. When the box body 212 slides relative to the reagent card 211, the rubber piston 24 moves towards the blind end to compress the gas in the gas source cavity and extrude it into the liquid cavity 2121.

[0123] Specifically, the box body 212 is provided with a sample adding port 2124, which can communicate with the liquid cavity 2121. The nucleic acid detection kit 21 further comprises an upper cover 2123 connected with the box body 212. The upper cover 2123 has a protrusion 2125 capable of sealing the sample adding port 2124. The quality control internal standard cavity 2126 has a first lyophilized ball inlet 21211 and a first lyophilized ball outlet 21212. The first lyophilized ball outlet 21212 communicates the liquid cavity 2121 and the quality control internal standard cavity 2126. The upper cover 2123 can also cover the first lyophilized ball inlet 21211.

[0124] Specifically, the first lyophilized ball 80 is an internal standard, which verifies the lysate stored in the liquid cavity 2121, the lysate stored in the nucleic acid amplification cavity 2111, the lyophilized ball system stored in the nucleic acid amplification cavity 2111, the heating module, the detection module, and the display module. The bottom of the quality control internal standard cavity 2126 is inclined at a certain angle with the horizontal plane, and the first lyophilized ball outlet 21212 is located at the lowest position of the inclined position. In this way, the first lyophilized ball 80 can move downward along the slope and enter the liquid cavity 2121 through the first lyophilized ball outlet 21212. In this embodiment, considering the light weight of the first lyophilized ball 80, a plastic ball 84 is pre-stored in the quality control internal standard cavity 2126. The plastic ball 84 can be a PP plastic ball. During the detection process, the plastic ball 84 can smoothly push the first lyophilized ball 80 into the liquid cavity 2121 under the action of its own gravity and the slope.

[0125] Specifically, the liquid cavity 2121 is provided with a second sealing film 26 at one end away from the sample inlet 2124. The liquid cavity 2121 can be connected with the puncture needle 25 to make one end of the puncture needle 25 pass through the second sealing film 26 and communicate with the liquid cavity 2121. The first lyophilized ball inlet 21211 is provided with a third sealing film 2127, and the first lyophilized ball outlet 21212 is provided with a fourth sealing film 2128. The second sealing film 26 and the third sealing film 2127 can be aluminum films. The wax column 2119, the sample inlet flow channel 2113, the main flow channel 2114, the waste gas flow channel 2115, the nucleic acid amplification cavity 2111, and the gas tail cavity 2112 are sealed by the first sealing film 2110, which can be an aluminum film.

[0126] More specifically, the fourth sealing film 2128 includes an integrally formed first part 21281, a second part 21282, and a third part 21283. The first part 21281, the second part 21282, and the third part 21283 are similar to a "concave" shape. The first part 21281 seals the first lyophilized ball outlet 21212, the second part 21282 seals the upper end of the liquid cavity 2121, and the third part 21283 is a pull ring structure that can extend out of the sample inlet 2124. When sample addition is needed, the fourth sealing film 2128 can be removed by holding the third part 21283 with the hand. In this embodiment, the fourth sealing film 2128 can be an aluminum-plastic composite film.

[0127] According to Figures 1-8 It can be seen that the nucleic acid detection kit 21 includes a box body 212 and a reagent card 211 connected by a connecting piece.

[0128] Specifically, the connecting piece of the connecting box body 212 and the reagent card 211 is a buckle assembly 213; it includes a buckle piece arranged on the reagent card 211 and a buckle 2132 arranged on the box body 212; wherein the buckle piece includes a buckle strip 2131, one end of the buckle strip 2131 is connected with the reagent card 211, the other end is a free end, and extends to the box body 212 along the buckle strip 2131, the buckle strip 2131 has a first cantilever 21311, the reagent card 211 has a second cantilever 2133, and the second cantilever 2133 is located above the first cantilever 21311; the buckle 2132 includes a first buckle corresponding to the first cantilever 21311 and a second buckle corresponding to the second cantilever 2133;

[0129] When the puncture needle 25 does not extend into the liquid cavity 2121, that is, the box body 212 does not slide downward relative to the reagent card 211, the first cantilever 21311 and the second cantilever 2133 are in abutment with the box body 212, ensuring that the reagent card 211 and the box body 212 do not displace each other;

[0130] When the puncture needle 25 extends into the liquid cavity 2121, that is, after the box body 212 slides relative to the reagent card 211, the first cantilever 21311 is coupled with the first buckle, and the second cantilever 2133 is coupled with the second buckle.

[0131] The above-mentioned microfluidic nucleic acid detection kit 21 can completely separate from the nucleic acid detection kit 21 in the laboratory, and the nucleic acid extraction is directly completed in the nucleic acid detection kit 21. At the same time, an internal air pressure balancing system is used in the nucleic acid detection kit 21 to store waste gas in the nucleic acid detection kit 21, so as to achieve fully closed sampling detection, completely eliminate the pollution risk of aerosol, and reduce the external power of the equipment by pushing the sample in the nucleic acid detection kit 21.

[0132] Embodiment two,

[0133] Referring to Figures 10 to 14 , a nucleic acid detection device is provided, which comprises a first shell 10, the first shell 10 has a shell body 11, the shell body 11 is provided with the microfluidic nucleic acid detection kit 21 provided in embodiment one, a first heating module and a first detection module. The nucleic acid detection device further comprises a display module arranged on the shell body 11, the display module comprises an LED lamp and has a display function, which is convenient for the operator to judge the detection result. In order to provide support, a mounting bottom plate 12 is arranged at the bottom of the shell body 11, and the shell body 11 is connected with the mounting bottom plate 12.

[0134] The upper end surface of the shell body 11 has an opening and a shell upper cover 13 rotationally connected with the shell body 11 and used for covering the opening, the nucleic acid detection kit 21 is inserted into the opening, the shell body 11 is provided with a mounting position for mounting a display module, and the display module is arranged on the mounting position.

[0135] The back surface of the shell body 11 is provided with a back shell 14, the mounting bottom plate 12 has a back shell mounting bottom groove 122, the shell body 11 has an upper groove for mounting the back shell 14, the back shell 14 is clamped between the back shell mounting bottom groove 122 and the upper groove, and the back shell 14 is provided with a debugging port 15, a first power socket 16 and a power switch 17 interface and the like. In order to realize automatic control, the shell body 11 is further provided with a power module, a data transmission and processing module and a control module. The nucleic acid detection device also has Bluetooth and WIFI functions, and can be remotely operated and the state, data and detection results of the nucleic acid detection device can be viewed in real time through a mobile terminal. The control principle of the electric type will not be described here, and the prior art can be referred to.

[0136] Reference Figure 13 The nucleic acid detection device includes a first circuit board 22 arranged on the mounting bottom plate 12 in the first shell 10 and a control unit arranged on the first circuit board 22, the nucleic acid detection kit 21 can be inserted into the opening on the shell body 11, and the control unit is electrically connected with the first heating module 3 to control the first heating module 3 to heat the nucleic acid amplification cavity 2111.

[0137] Reference Figure 12 、 13 and 14, the first heating module 3 includes a heating block 31 closely surrounding the reaction cavity, a first heating sheet 32 arranged at the bottom of the heating block 31 and in contact with the heating block 31 for heat conduction, and a temperature sensor adjacent to the first heating sheet 32, the temperature sensor is electrically connected with the control unit, and a heat dissipation space is formed between the first heating module 3 and the first circuit board 22.

[0138] Specifically, the heating block 31 is provided with a heating groove 311 matched with the nucleic acid amplification cavity 2111 arranged in the reagent card 211, the nucleic acid amplification cavity 2111 can extend into the heating groove 311, the heating block 31 is provided with a first hole 312 and a second hole arranged oppositely, the first hole 312 and the second hole both penetrate the heating groove 311, and the nucleic acid amplification cavity 2111 is arranged between the first hole 312 and the second hole;

[0139] In the embodiment, the first holes 312 include three, namely the first hole I, the first hole II and the first hole III. Obviously, the corresponding second holes also include three, namely the second hole I corresponding to the first hole I, the second hole II corresponding to the first hole II and the second hole III corresponding to the first hole III. The nucleic acid amplification cavity one is located between the first hole I and the second hole I, the nucleic acid amplification cavity two is located between the first hole II and the second hole II, and the nucleic acid amplification cavity three is located between the first hole III and the second hole III.

[0140] The first detection module can be designed according to different detection methods. The nucleic acid detection method includes hybridization fluorescence, isothermal amplification, real-time fluorescent PCR, high-resolution melting curve, electrochemical nucleic acid aptamer, etc. The corresponding detection module is designed according to the detection method.

[0141] In the embodiment, the first detection module includes the light source 41 and the light sensor 42 electrically connected with the first circuit board 22. The light source 41 and the light sensor 42 are both arranged outside the heating block 31, and the light source 41 is attached to the first hole 312, and the light sensor 42 is attached to the second hole. Specifically, the first heating module 3 heats the nucleic acid amplification cavity one, the nucleic acid amplification cavity two and the nucleic acid amplification cavity three. At the same time, the light source 41 reaches the light sensor 42 through the nucleic acid amplification cavity one, the nucleic acid amplification cavity two and the nucleic acid amplification cavity three respectively, and the light sensor 42 receives the difference of the change of the light signal.

[0142] In the embodiment, the first circuit board 22 is installed on the circuit board mounting seat 121, and the circuit board mounting seat 121 is arranged in the mounting bottom plate 12. The first circuit board 22 is provided with a circuit board cover plate 123. The first circuit board 22 includes a processing unit. The processing unit is used for processing and analyzing the light signal collected by the first heating module 3, and then feeding back the analysis result to the display module.

[0143] Specifically, the processing unit includes a data acquisition circuit, an ADC chip and a processor. The data acquisition circuit collects the signal of the light sensor. After analog-digital conversion by the ADC chip, the digital signal is processed and analyzed by the processor, and then the analysis result is fed back to the display module. That is, the processing unit first collects the light signal on the light sensor, which can be fed back to the LED lamp. After analog-digital conversion of the light signal change signal, the digital signal is processed and analyzed by the processor, and the result is fed back to the display module.

[0144] The specific operation steps are as follows:

[0145] Sample addition

[0146] Open the upper cover 2123 of the box body 212, and remove the aluminum composite film. At this time, the first freeze-dried ball 80 falls into the liquid cavity 2121 under the pushing action of the plastic ball 84. The throat swab is swished in the lysis solution for ten or so circles. In the process of continuously swishing the throat swab, the first freeze-dried ball 80 is also uniformly mixed in the lysis solution. After swishing, the upper cover 2123 is closed.

[0147] 2. Lysis

[0148] The reagent card 211 of the nucleic acid detection kit 21 is inserted into the opening on the shell body 11. The box body 212 is lightly pressed downward to be coupled with the first cantilever and the first clamping opening, and coupled with the second cantilever and the second clamping opening. At this time, the puncture needle 25 pierces the second sealing film 26, and at the same time, the rubber piston 24 pushes the gas in the gas cavity 2122 to the liquid cavity 2121, so that the pressure in the liquid cavity 2121 is increased. In this embodiment, considering the consistency of the liquid entering the nucleic acid amplification cavity 2111, the atmospheric pressure in the liquid cavity 2121 is 0.137 MPa at this time. The sample is lysed in the lysis solution for 10 min.

[0149] 3. Amplification

[0150] After the sample is lysed in the lysis solution for 10 min, the first heating module 3 heats the wax column 2111. At this time, the wax column 2119 melts, so that all the flow channels are conducted. Under the action of the pressure difference, the liquid enters the main flow channel 2114 through the sample inlet flow channel 2113, and then enters the nucleic acid amplification cavity one, the nucleic acid amplification cavity two and the nucleic acid amplification cavity three through the branch flow channels, respectively. The nucleic acid amplification cavity one is pre-embedded with the freeze-dried ball one 81, the nucleic acid amplification cavity two is pre-embedded with the freeze-dried ball two 82, and the nucleic acid amplification cavity three is pre-embedded with the freeze-dried ball three 83. In this process, the first heating module 3 heats and amplifies the nucleic acid amplification cavity one, the nucleic acid amplification cavity two and the nucleic acid amplification cavity three.

[0151] 4. Detection

[0152] After amplification, the amplification results of the nucleic acid amplification cavity one, the nucleic acid amplification cavity two and the nucleic acid amplification cavity three are detected.

[0153] The device is simple and convenient to operate. The nucleic acid detection kit 21 extracts the sample through the sample inlet in the box, has no gas outlet, is completely closed and has no pollution detection, and is completely separated from the laboratory use. At the same time, the nucleic acid detection device uses the card to push the sample, without external power. The nucleic acid detection device is simple, and the complexity of the detection equipment is simplified.

[0154] Example three,

[0155] Reference Figures 15-19The embodiment provides an integrated nucleic acid detection device, which comprises the microfluidic nucleic acid detection kit 21 provided in the first embodiment and a second shell for accommodating the nucleic acid detection kit 21. The second shell can be integrally formed. The second shell can also comprise a front cover 51 and a rear cover 52 for clamping the nucleic acid detection kit 21. The nucleic acid detection kit 21 is arranged between the front cover 51 and the rear cover 52. The integrated nucleic acid detection device further comprises a second heating module 6 for heating the nucleic acid amplification cavity 2111; a second detection module arranged between the front cover 51 and the rear cover 52 and used for detecting changes in an optical signal in the nucleic acid amplification cavity 2111. The optical signal comprises one or more of color, brightness or fluorescence saturation.

[0156] The nucleic acid detection kit 21 and the detection device of the integrated nucleic acid detection device are integrated, so that the kit and the detection device are no longer separated, forming an integrated and disposable nucleic acid detection kit 21, which is an indivisible whole. One person uses one kit and discards it. The detection equipment is abandoned, and the pollution of the detection device to the card is prevented. Meanwhile, the integrated nucleic acid detection device is a nucleic acid detection device completely separated from a laboratory, and achieves full-closed sample injection detection, completely eliminating the risk of aerosol pollution.

[0157] Specifically, as Figure 17 and Figure 18As shown, the front cover 51 and the rear cover 52 can be snap-fit connected, wherein the front cover 51 comprises a first bottom surface 514, a first side surface 1515, a second side surface 1516 and a third side surface 1517 integrally formed, and the first bottom surface 514, the first side surface 1515, the second side surface 1516 and the third side surface 1517 are enclosed in a "concave" shape, the first side surface 1515 and the second side surface 1516 are symmetrically arranged on both sides of the first bottom surface 514, and a plurality of fixing columns 512 are symmetrically arranged on the first side surface 1515 and the second side surface 1516 for connecting the rear cover 52; the rear cover 52 is in a "concave" shape and comprises a second bottom surface 522, a first side surface 1523, a second side surface 1524 and a third side surface 1525 integrally formed, wherein the first side surface 1523 and the second side surface 1524 are symmetrically arranged on both sides of the second bottom surface 522, and a plurality of fixing grooves 521 matching the fixing columns 512 are symmetrically arranged on the first side surface 1523 and the second side surface 1524, the front cover 51 and the rear cover 52 are snap-fit connected together through the snap-fit of the fixing columns 512 and the fixing grooves 521, wherein the first side surface 1515 and the first side surface 1523 are butted against each other to form a first side edge of the shell, the second side surface 1516 and the second side surface 1524 are butted against each other to form a second side edge of the shell, and the third side surface 1517 and the third side surface 1525 are butted against each other to form a bottom plate of the shell, and the cavity formed after the butting of the front cover 51 and the rear cover 52 can accommodate the microfluidic nucleic acid detection kit 21 in the embodiment, and the box body 212 of the nucleic acid detection kit 21 is exposed to the cavity enclosed by the front cover 51 and the rear cover 52.

[0158] A second circuit board 62 is further arranged between the front cover 51 and the rear cover 52, one end of the second circuit board 62 is inserted into the box body 212 of the nucleic acid detection kit 21, and the other end thereof is exposed to the box body 212 and flush with the reagent card 211, the second circuit board 62 is provided with a second LED lamp 72, and a control unit is arranged on the second circuit board 62, the control unit is electrically connected with the second heating module 6 to control the second heating module 6 to heat the nucleic acid amplification cavity 2111, and a second power socket 621 is further arranged on the second circuit board 62, and the side edges of the front cover 51 and the rear cover 52 are both provided with notches for exposing the second power socket 621. The second circuit board 62 is provided with a data communication unit, and the data communication unit is used for transmitting detection data to a wireless terminal, wherein the wireless terminal includes but is not limited to a mobile phone terminal, a computer or a network server. In the embodiment, the second circuit board 62 is installed on the rear cover 52, and the second circuit board 62 comprises a processing unit, and the processing unit is used for processing and analyzing the optical signal collected by the detection module to obtain detection data.

[0159] Specifically, the processing unit comprises a data acquisition circuit, an ADC chip and a processor, the data acquisition circuit acquires the signal of the temperature sensor 63, the signal is converted into digital signal by the ADC chip, and then the analysis result is fed back to the processing unit.

[0160] As Figure 19 shown, the second heating module 6 comprises a second heating sheet 61, wherein the second heating sheet 61 has two groups and is oppositely arranged, one end of the two groups of second heating sheet 61 is connected through a connecting piece, the other end of the two groups of second heating sheet 61 is open, the two groups of second heating sheet 61 enclose the nucleic acid amplification cavity 2111, three third holes 64 are arranged on one of the second heating sheet 61, and the other second heating sheet is provided with fourth holes corresponding to the third holes 64, and the third holes and the fourth holes each have three groups, which respectively surround the nucleic acid amplification cavity one, the nucleic acid amplification cavity two and the nucleic acid amplification cavity three.

[0161] The second detection module can be designed according to different detection methods, and the nucleic acid detection method includes hybridization fluorescence, isothermal amplification, real-time fluorescent PCR, high-resolution melting curve, electrochemical nucleic acid aptamer and the like. The corresponding detection module is designed according to the detection method.

[0162] In the embodiment, the second detection module comprises a second LED lamp 72 and a light guide column 71 electrically connected with the second circuit board 62, and the second LED lamp 72 and the light guide column 71 are respectively arranged on both sides of the reagent card 211. Specifically, the second heating module 6 heats the nucleic acid amplification cavity one, the nucleic acid amplification cavity two and the nucleic acid amplification cavity three, at the same time, the second LED lamp 72 respectively passes through the nucleic acid amplification cavity one, the nucleic acid amplification cavity two and the nucleic acid amplification cavity three to reach the light guide column 71, and the user observes the color change of the light guide column 71.

[0163] The above embodiments only illustrate the basic principles and characteristics of the present application, and the present application is not limited to the above embodiments, and various changes and changes can be made without departing from the spirit and scope of the present application, and these changes and changes all fall within the scope of the present application. The scope of protection of the present application is defined by the appended claims and their equivalents.

Claims

1. A microfluidic nucleic acid detection kit, comprising an interconnected housing and reagent cards, characterized in that, The housing is slidable relative to the reagent card to control the operation of the reagent kit, and further includes several cavities and several channels for communicating with the cavities. The cavities and channels form a closed system. The channels include a gas source channel, a sample inlet channel, and a waste gas channel. The cavities include at least: An air source chamber is located inside the box and its volume changes can control the air pressure of the sealed system. A liquid chamber, disposed within the box, is used to inject liquid reagents; the liquid chamber and the gas source chamber are connected via a gas source flow channel. A nucleic acid amplification chamber is disposed on the reagent card, and the nucleic acid amplification chamber is connected to the liquid chamber through a sample inlet channel; A gas tail cavity is provided on the reagent card, and the gas tail cavity is connected to the nucleic acid amplification cavity through a waste gas flow channel; A first sealing membrane is disposed on the reagent card, and the first sealing membrane seals the sample inlet channel, the waste gas channel and the gas tail cavity; Before the cartridge slides relative to the reagent card, the air pressure in the gas source chamber and the air pressure in the gas tail chamber are the same, and the liquid in the liquid chamber will not flow into the nucleic acid amplification chamber. When the cartridge slides relative to the reagent card, the gas source chamber is compressed, and the air pressure in the gas source chamber increases, thereby pushing the liquid reagent in the liquid chamber to be injected into the nucleic acid amplification chamber through the sample inlet channel. The flow of the liquid reagent pushes the gas in the nucleic acid amplification chamber to be injected into the gas tail chamber.

2. The microfluidic nucleic acid detection kit according to claim 1, characterized in that, An overflow valve is installed at the waste gas passage. The overflow valve is used to block the flow of liquid while allowing gas to be discharged.

3. The microfluidic nucleic acid detection kit according to claim 1, characterized in that, The plurality of cavities also includes a bubble storage cavity, which is disposed between the waste gas flow channel and the nucleic acid amplification cavity, and the bubble storage cavity is connected to the nucleic acid amplification cavity.

4. The microfluidic nucleic acid detection kit according to claim 2, characterized in that, The box body and the reagent card are connected by a snap-fit ​​assembly, which includes a snap-fit ​​element disposed on the reagent card and a snap-fit ​​buckle disposed on the box body; wherein... The fastener includes a fastening strip, one end of which is connected to the reagent card, and the other end of which is a free end that extends along the fastening strip toward the box body. The fastening strip has a first cantilever, and the reagent card has a second cantilever. The second cantilever is located above the first cantilever. The latch includes a first latch corresponding to the first cantilever and a second latch corresponding to the second cantilever.

5. The microfluidic nucleic acid detection kit according to claim 4, characterized in that, Before the box body slides relative to the reagent card, both the first cantilever and the second cantilever are in contact with the box body, the air pressure of the gas source chamber and the gas tail chamber are the same, and the liquid in the liquid chamber will not flow into the nucleic acid amplification chamber; When the liquid reagent in the liquid chamber reaches the overflow valve, the liquid reagent stops flowing, thereby injecting the liquid reagent into the nucleic acid amplification chamber.

6. The microfluidic nucleic acid detection kit according to claim 5, characterized in that, The nucleic acid amplification chamber contains solid reagents required for nucleic acid amplification, including dry powder reagents, lyophilized reagent bulbs, or quality control internal standard reagents; the liquid chamber contains nucleic acid lysis extract and provides space for adding samples, and the volume of the nucleic acid lysis extract is 100μL~5000μL.

7. The microfluidic nucleic acid detection kit according to claim 1, characterized in that, A main channel is provided between the nucleic acid amplification chamber and the sample inlet channel, and a wax column is provided between the sample inlet channel and the main channel.

8. The microfluidic nucleic acid detection kit according to claim 6, characterized in that, The plurality of cavities also includes a quality control internal standard cavity disposed within the box, wherein a first lyophilized bulb is placed in the quality control internal standard cavity, and the first lyophilized bulb contains the reagent components required for internal quality control.

9. The microfluidic nucleic acid detection kit according to claim 1, characterized in that, The nucleic acid amplification chamber contains a second lyophilized bulb or lyophilized powder, which contains the reagent components required for nucleic acid amplification.

10. The microfluidic nucleic acid detection kit according to claim 8, characterized in that, The bottom of the quality control internal standard cavity is inclined at a certain angle to the horizontal plane, and the outlet of the first freeze-dried bulb is located at the lowest position of the bottom of the quality control internal standard cavity.

11. The microfluidic nucleic acid detection kit according to claim 6, characterized in that, The sample inlet channel has a puncture needle at one end near the liquid chamber. The puncture needle can be inserted into the liquid chamber so that the puncture needle extends into the liquid chamber, thereby connecting the sample inlet channel with the liquid chamber.

12. The microfluidic nucleic acid detection kit according to claim 1, characterized in that, The reagent card has a piston post, and a rubber piston is provided at one end of the piston post. The air source chamber is in the shape of a blind hole on the box body. One end of the piston rod with the rubber piston extends into the blind end of the air source chamber through the open end of the air source chamber. The air source flow channel is located near the blind end of the air source chamber. Before the box body slides relative to the reagent card, the rubber piston is lower than the gas source flow channel in the gas source chamber; When the box slides relative to the reagent card, the rubber piston moves toward the blind end to compress the gas in the gas source chamber and squeeze it into the liquid chamber.

13. The microfluidic nucleic acid detection kit according to claim 8, characterized in that, The box body has a sample inlet that can communicate with the liquid chamber. The liquid chamber has a sample inlet at the end away from the sample inlet channel. The quality control internal standard chamber has a first lyophilized bulb inlet and a first lyophilized bulb outlet. The first lyophilized bulb outlet communicates with the liquid chamber and the quality control internal standard chamber.

14. The microfluidic nucleic acid detection kit according to claim 13, characterized in that, The box body also includes a top cover, which has a protrusion capable of sealing the sample dispensing port and the outlet of the first lyophilized bulb, and the top cover is capable of covering the inlet of the first lyophilized bulb.

15. The microfluidic nucleic acid detection kit according to claim 13, characterized in that, A second sealing membrane is provided at one end of the liquid chamber away from the sample application port. The liquid chamber can be inserted into a puncture needle so that one end of the puncture needle passes through the second sealing membrane and communicates with the liquid chamber. A third sealing membrane is provided at the inlet of the first lyophilized bulb, and a fourth sealing membrane is provided at the outlet of the first lyophilized bulb. The fourth sealing membrane includes an integrally formed first part, a second part, and a third part. The first part seals the outlet of the first freeze-dried bulb, the second part seals the liquid cavity, and the third part is a pull ring structure that can extend out of the sample dispensing port.

16. A detection device, characterized in that, The microfluidic nucleic acid detection kit according to any one of claims 1-15 further includes a first housing, a power module, and a display module, wherein the nucleic acid detection kit is detachably plugged into the first housing, and the detection device further includes: The first heating module is used to heat the nucleic acid amplification chamber; The first detection module, which is disposed within the first housing, is used to detect changes in the optical signal within the nucleic acid amplification chamber. The optical signal includes one or more of the following: color, brightness, fluorescence, or saturation signals.

17. The detection device according to claim 16, characterized in that, It also includes a first circuit board disposed within the first housing and a control unit disposed on the first circuit board, the control unit being electrically connected to the first heating module to control the first heating module to heat the nucleic acid amplification chamber.

18. The detection device according to claim 17, characterized in that, The first heating module includes a heating block that tightly surrounds the nucleic acid amplification cavity, a first heating element disposed at the bottom of the heating block and in contact with the heating block for heat conduction, and a temperature sensor adjacent to the first heating element. The temperature sensor is electrically connected to the control unit, and a heat dissipation space is formed between the first heating module and the first circuit board.

19. The detection device according to claim 18, characterized in that, The heating block has a heating groove and a first hole and a second hole arranged opposite to each other. The first hole and the second hole both penetrate the heating groove. The nucleic acid amplification cavity can extend into the heating groove and is located between the first hole and the second hole.

20. The detection device according to claim 19, characterized in that, The first detection module includes a light source and a light sensor electrically connected to the first circuit board. The light source and the light sensor are both disposed on the outside of the heating block, and the light source is attached to the first hole, while the light sensor is attached to the second hole.

21. The detection device according to claim 20, characterized in that, The first circuit board has a processing unit, which is used to process and analyze the optical signal collected by the detection module, and then feed the analysis result back to the display module.

22. The detection device according to claim 21, characterized in that, The processing unit includes a data acquisition circuit, an ADC chip, and a processor. The data acquisition circuit acquires signals from the optical sensor, which are then converted from analog to digital by the ADC chip and sent to the processor for digital signal processing and analysis to obtain detection data.

23. The detection device according to claim 22, characterized in that, The first circuit board has a data communication unit, which is used to transmit the detection data to a wireless terminal, the wireless terminal including one or more of a mobile phone terminal, a computer or a network server.

24. The detection device according to claim 23, characterized in that, The first housing includes a housing body with an opening at the upper end and a housing cover rotatably connected to the housing body and used to close the opening. The nucleic acid detection kit is detachably inserted into the opening. The housing body is provided with a mounting position for installing the display module, and the display module is disposed in the mounting position.

25. An integrated detection device, characterized in that, The integrated detection device includes the microfluidic nucleic acid detection kit according to any one of claims 1-15 and a second housing for holding the microfluidic nucleic acid detection kit, and further includes: The second heating module is used to heat the nucleic acid amplification chamber; The second detection module, disposed within the second housing, is used to detect changes in the optical signal within the nucleic acid amplification chamber. The optical signal includes one or more of the following: color, brightness, fluorescence, or saturation.

26. The integrated detection device according to claim 25, characterized in that, The housing includes a detachably connected front cover and a rear cover, which cooperate to hold the microfluidic nucleic acid detection kit, and the second detection module is disposed between the front cover and the rear cover.

27. The integrated detection device according to claim 26, characterized in that, It also includes a second circuit board disposed between the front cover and the rear cover, and a control unit disposed on the second circuit board. The control unit is electrically connected to the second heating module to control the second heating module to heat the nucleic acid amplification chamber. The second circuit board has a second power socket.

28. The integrated detection device according to claim 27, characterized in that, The second heating module includes a second heating element that tightly surrounds the nucleic acid amplification chamber and a temperature sensor disposed on the inner surface of the second heating element and in contact with the reagent card. The temperature sensor is electrically connected to the control unit.

29. The integrated detection device according to claim 27, characterized in that, The second detection module includes a light source and a light guide column electrically connected to the second circuit board, with the light source and the light guide column located on opposite sides of the reagent card.

30. The integrated detection device according to claim 29, characterized in that, The second circuit board has a processing unit, which is used to process and analyze the optical signal collected by the second detection module to obtain detection data.

31. The integrated detection device according to claim 30, characterized in that, The processing unit includes a data acquisition circuit, an ADC chip, and a processor. The data acquisition circuit acquires signals from the optical sensor, which are then converted from analog to digital by the ADC chip and sent to the processor for digital signal processing and analysis to obtain detection data.

32. The integrated detection device according to claim 31, characterized in that, The second circuit board has a data communication unit, which is used to transmit the detection data to a wireless terminal, including one or more of a mobile phone terminal, a computer, or a network server.

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