Portable pine wood nematode DNA detection card and detection module

Abstract

The utility model relates to a portable bursaphelenchus xylophilus DNA detection card and a detection module, and belongs to the technical field of bursaphelenchus xylophilus detection, the portable bursaphelenchus xylophilus DNA detection card comprises a detection card body, the detection card body is provided with a sample liquid injection port, a remelting unit, an amplification reaction unit, a dilution unit and a detection unit, the sample liquid injection port is connected with the remelting unit, and the amplification reaction unit is connected with the dilution unit. The remelting unit is connected with the amplification reaction unit, the amplification reaction unit is connected with the dilution unit, the diluent unit is connected with the detection unit, and the detection card body is provided with a control assembly used for controlling liquid to flow from the sample liquid injection port to the detection unit. The pine wood nematode detection device has the advantages that sampling personnel can detect pine wood nematode on a sampling site, and a detection result can be accurately obtained.

Description

Technical Field

[0001] This application relates to the technical field of pine wood nematode detection, and particularly to a portable pine wood nematode DNA detection card and a detection module. Background Art

[0002] Pine wilt disease is a worldwide forestry disease caused by the infection of pine wood nematodes, which can damage more than 70 coniferous trees including 57 pine tree species. In recent decades, the cumulative number of pine trees lost due to pine wilt disease in China has still reached billions, causing economic losses of up to hundreds of billions of yuan, and the disease is still spreading rapidly in China. When detecting whether a pine tree is infected with pine wood nematodes, the Baermann funnel method is generally used. The tester samples the nematode fluid exuded from the pine wood and observes it under a microscope. However, since there are many types of nematodes in pine trees, not all nematode species are harmful to pine tree species. Many nematodes have similar appearances but different characteristics. When detecting by the Baermann funnel method under a microscope, it not only requires high professional knowledge for the discriminator, but there may be misjudgments. Moreover, it usually takes 8 - 10 hours to exude nematode fluid from the pine wood, and the overall detection time is long and the difficulty is high. The existing detection methods also use PCR equipment for DNA detection. The DNA detection method has high precision, but the PCR equipment is relatively large in volume and is usually placed in a laboratory, and it is impossible to immediately obtain the detection result after on-site sampling, resulting in a long detection cycle for pine wood nematodes. Summary of the Utility Model

[0003] In order to enable the sampler to detect pine wood nematodes at the sampling site and accurately obtain the detection result, this application provides a portable pine wood nematode DNA detection card and a detection module.

[0004] A portable pine wood nematode DNA detection card provided by this application adopts the following technical solution:

[0005] A portable pine wood nematode DNA detection card includes a detection card body. A sample liquid injection port, a re - melting unit, an amplification reaction unit, a dilution unit, and a detection unit are arranged on the detection card body. The sample liquid injection port is connected to the re - melting unit, the re - melting unit is connected to the amplification reaction unit, the amplification reaction unit is connected to the dilution unit, the dilution unit is connected to the detection unit, and a control component for controlling the flow of liquid from the sample liquid injection port in the direction of the detection unit is arranged on the detection card body.

[0006] By adopting the above technical solution, the pine wood nematode liquid or pine wood chips are heated to 95°C / 5 minutes with special reagents on site for lysis, and then the lysis-completed sample liquid is injected into the sample liquid injection port, and the sample liquid is injected into the remelting unit for remelting through the control component, and then flows into the amplification reaction unit for amplification reaction. After heating the sample liquid in the amplification reaction unit, the amplified sample liquid is discharged into the dilution unit for dilution, and finally the diluted sample liquid is discharged into the detection unit for detection, and the detection result can be obtained by observing the information displayed by the detection unit. The sampling personnel can perform pine wood nematode detection at the sampling site and accurately obtain the detection results, and the operation is simple and convenient for detection. Moreover, this disposable detection card is easy to carry, and no transfer is required during the sample liquid detection process, which reduces the contamination of the sample liquid.

[0007] Optionally, the remelting unit includes a remelting liquid tank opened on the detection card body, the remelting liquid tank is filled with remelting liquid, and the notch of the remelting liquid tank is sealed by a sealing film.

[0008] Optionally, the amplification reaction unit includes an amplification reaction groove opened on the detection card body, the amplification reaction groove is filled with freeze-dried balls, the groove of the amplification reaction groove is sealed by a sealing film, and a first opening and closing member is provided at the discharge of the amplification reaction groove.

[0009] By adopting the above technical solution, when the amplification reaction is performed, the first opening and closing member closes the drainage of the amplification tank to prevent the sample liquid from flowing into the dilution unit, and the control component restricts the sample liquid from flowing back into the remelting unit, so that the sample liquid and the freeze-dried ball can fully perform the amplification reaction. When the amplification reaction is completed, the first opening and closing member opens the drainage of the amplification tank, and the amplified sample liquid can be discharged into the dilution unit for dilution.

[0010] Optionally, the first opening and closing member includes a first paraffin wax, and the melting point of the first paraffin wax is higher than the amplification reaction temperature.

[0011] By adopting the above technical solution, when performing an amplification reaction, the sample liquid, remelting liquid and freeze-dried balls are heated to 40 degrees and maintained for 20 minutes. At this time, the first paraffin will not melt. After the amplification reaction is completed, the heating temperature is increased again to melt the first paraffin, so that the amplified sample liquid can be smoothly discharged into the dilution unit for dilution. Due to the chemical inertness of the first paraffin, it will not mix with the sample liquid after melting, which can avoid interference with the detection results.

[0012] Optionally, the dilution unit comprises a dilution liquid tank provided on the detection card body, the dilution liquid tank is filled with dilution liquid, a slot of the dilution liquid tank is sealed by a sealing film, and a second opening and closing member is provided at the discharge of the dilution liquid tank.

[0013] By adopting the above technical solution, before the dilution is completed, the second opening and closing member closes the liquid discharge part of the dilution liquid tank to prevent the sample liquid from flowing into the detection unit. After the dilution is completed, the second opening and closing member opens the liquid discharge part of the dilution liquid tank. At this time, the diluted sample liquid can be discharged into the detection unit for detection.

[0014] Optionally, the second opening and closing member includes second paraffin wax, and the melting point of the second paraffin wax is higher than that of the first paraffin wax.

[0015] By adopting the above technical solution, when the heating temperature reaches the melting point of the first paraffin wax, the second paraffin wax will not melt at this time. The amplified sample liquid can flow smoothly into the dilution liquid tank but will not flow into the detection unit. After the dilution is completed, raising the heating temperature again can melt the second paraffin wax, so that the liquid discharge part of the dilution tank is opened, and the sample liquid can flow smoothly into the detection unit for detection.

[0016] Optionally, the detection unit includes a detection groove opened on the detection card body, a test strip is inserted into the detection groove, and the notch of the detection port is sealed by a sealing film.

[0017] By adopting the above technical solution, after the diluted sample liquid flows into the detection groove, the test strip absorbs the sample liquid. By checking the information displayed on the test strip, it can be judged whether there are harmful pine wood nematodes in the sample liquid.

[0018] Optionally, the control component includes a one-way pressing cap, the one-way pressing cap is detachably arranged on the sample liquid injection port, and first one-way valves are arranged at the connection between the sample liquid injection port and the re-melting unit and at the connection between the re-melting unit and the amplification reaction unit.

[0019] By adopting the above technical solution, when injecting the sample liquid into the detection card body, first remove the one-way pressing cap from the sample liquid injection port, then inject the sample liquid into the sample liquid injection port, and then reinstall the one-way pressing cap. By squeezing the one-way pressing cap, the sample liquid can be injected into the re-melting unit for re-melting, and then continuously pressing the one-way pressing cap can discharge the re-melted sample liquid into the amplification unit. And by pressing the one-way pressing cap multiple times, inflation can be continuously carried out to push the diluted sample liquid into the detection unit for detection.

[0020] A portable pine wood nematode DNA detection module provided by the present application adopts the following technical solution:

[0021] A portable DNA detection module for Bursaphelenchus xylophilus, comprising the above-mentioned portable DNA detection card for Bursaphelenchus xylophilus and a heating unit. The heating unit includes a housing, a heating sheet is arranged inside the housing, a control module for controlling the heating sheet to heat at different temperatures is arranged inside the housing, a power module is arranged inside the housing, and the power module is connected to the control module.

[0022] By adopting the above technical solution, the heating unit can provide heating at different temperatures for the detection card body, and the power module provides power, without the need for an external power supply, which is convenient for the detection personnel to carry and convenient for heating operation at the sampling site.

[0023] Optionally, at least two slots are opened inside the housing. The number of heating sheets is the same as and corresponds to the number of slots. The area of the slot facing the heating sheet is the heating area. The detection card body can be inserted into the slot. When the detection card body is inserted properly, the amplification reaction unit and the dilution unit are exactly located in the heating area. At least two operation ports are opened on the housing, and the operation ports correspond to and communicate with the slots one by one.

[0024] By adopting the above technical solution, the setting of multiple slots and multiple heating sheets can realize heating of multiple detection card bodies simultaneously, improve the detection efficiency, and the detection card body can be fixed directly after being inserted into the slot. The sample liquid can be injected into the detection body through the operation port, which is convenient for operation.

[0025] In summary, the present application includes at least one of the following beneficial technical effects:

[0026] 1. It enables the sampling personnel to perform the detection of Bursaphelenchus xylophilus at the sampling site, accurately obtain the detection result, and the operation is simple and convenient for detection.

[0027] 2. This disposable detection card is convenient to carry, and there is no need for transfer during the detection process of the sample liquid, reducing the situation of sample liquid being contaminated.

[0028] 3. The heating unit can provide heating at different temperatures for the detection card body, and the power module provides power, without the need for an external power supply, which is convenient for the detection personnel to carry and convenient for heating operation at the sampling site. Description of the Drawings

[0029] Figure 1 It is the overall structural schematic diagram of the embodiment of the present application.

[0030] Figure 2 It is the structural schematic diagram of the embodiment of the present application for showing the heating unit and the one-way pressing cap.

[0031] Figure 3It is a schematic structural diagram of the heating block used in the embodiments of the present application.

[0032] Explanation of reference numerals: 1. Detection card body; 11. Sample liquid injection port; 12. Connection groove; 2. Re-melting unit; 21. Re-melting liquid tank; 22. Re-melting liquid; 3. Amplification reaction unit; 31. Amplification reaction tank; 32. Freeze-dried ball; 33. First opening and closing member; 331. First paraffin wax; 4. Dilution unit; 41. Dilution liquid tank; 42. Dilution liquid; 43. Second opening and closing member; 431. Second paraffin wax; 5. Detection unit; 51. Detection tank; 52. Test strip; 6. Control component; 61. One-way pressing cap; 62. One-way valve; 7. Heating unit; 71. Housing; 711. Slot; 712. Operation port; 72. Heating sheet; 73. Power module; 74. HMI; 75. Control module Detailed implementation manners

[0033] The following will Figures 1-3 further elaborate on the present application in detail.

[0034] The embodiments of the present application disclose a portable DNA detection card for Bursaphelenchus xylophilus.

[0035] As Figure 1 and Figure 2 , the portable DNA detection card for Bursaphelenchus xylophilus includes a detection card body 1, and a sample liquid injection port 11, a re-melting unit 2, an amplification reaction unit 3, a dilution unit 4, and a detection unit 5 are arranged on the detection card body 1;

[0036] The re-melting unit 2 includes a re-melting liquid tank 21 opened on the detection card body 1, and a re-melting liquid 22 is pre-filled in the re-melting liquid tank 21. The re-melting liquid 22 is enzyme-free water, and the mouth of the re-melting liquid tank 21 is sealed with a sealing film. In other embodiments, the re-melting unit 2 can also be a convex housing filled with the re-melting liquid 22, and the convex housing is fixed on the detection card body 1.

[0037] The amplification reaction unit 3 includes an amplification reaction tank 31 opened on the detection card body 1, and a freeze-dried ball 32 is pre-filled in the amplification reaction tank 31. The freeze-dried ball 32 is a solid ball obtained by freeze-drying a solution containing Taq DNA polymerase, specific primers, probes, and dNTPs. A first opening and closing member 33 is arranged at the liquid discharge place of the amplification reaction tank 31, and the mouth of the amplification reaction tank 31 is sealed with a sealing film. In other embodiments, the amplification reaction unit 3 can also be a convex housing filled with the freeze-dried ball 32, and the convex housing is fixed on the detection card body 1.

[0038] The dilution unit 4 includes a dilution liquid tank 41 opened on the detection card body 1. The dilution liquid tank 41 is pre-filled with a dilution liquid 42, and the dilution liquid 42 is enzyme-free water. A second opening and closing member 43 is provided at the liquid discharge place of the dilution liquid tank 41. The mouth of the dilution liquid tank 41 is sealed with a sealing film. In other embodiments, the dilution unit 4 can also be a convex shell filled with the dilution liquid 42, and the convex shell is fixed on the detection card body 1.

[0039] The detection unit 5 includes a detection tank 51 opened on the detection card body 1. A test strip 52 is inserted into the detection tank 51. The mouth of the detection tank 51 is sealed with a sealing film, and at the same time, the sealing film fixes the test strip 52. In other embodiments, the detection unit 5 can also be a convex shell, and the convex shell is fixed on the detection card body 1, and one end of the test strip 52 is inserted into the convex shell.

[0040] The sample liquid injection port 11, the reconstitution liquid tank 21, the amplification reaction tank 31, and the dilution liquid tank 41 are arranged along the length direction of the detection card body 1 and are located on one side of the detection card body 1. The detection tank 51 and the test strip 52 are arranged on the other side of the detection card body 1. The sample liquid injection port 11 and the reconstitution liquid tank 21 are connected through a connection groove 12 opened on the detection card body 1. The reconstitution liquid tank 21 and the amplification reaction tank 31 are connected through a connection groove 12 opened on the detection card body 1. The amplification reaction tank 31 and the dilution liquid tank 41 are connected through a connection groove 12 opened on the detection card body 1. The dilution liquid tank 41 and the detection tank 51 are connected through a connection groove 12 opened on the detection card body 1. The first opening and closing member 33 is pre-fixed in the connection groove 12 between the amplification reaction tank 31 and the dilution liquid tank 41. The second opening and closing member 43 is pre-fixed in the connection groove 12 between the dilution liquid tank 41 and the detection tank 51. All the connection grooves 12 are sealed with a sealing film. In other embodiments, connection can also be achieved by externally installing connecting pipes or opening channels inside the detection card body 1.

[0041] A control component 6 for controlling the flow of the sample liquid from the sample liquid injection port 11 towards the detection tank 51 is further provided on the detection card body 1. The control component 6 includes a one-way pressing cap 61. The one-way pressing cap 61 is sleeved on the sample liquid injection port 11. The one-way pressing cap 61 is a rubber sleeve, and a one-way air outlet valve and a one-way air inlet valve are provided on the rubber sleeve. The inside of the rubber sleeve is a cavity structure. A one-way valve 62 is installed in the connection groove 12 between the sample liquid injection port 11 and the reconstitution liquid tank 21. A one-way valve 62 is also installed in the communication groove between the reconstitution liquid tank 21 and the amplification reaction tank 31. The opening pressure of the one-way valve 62 between the sample liquid injection port 11 and the reconstitution liquid tank 21 is less than the opening pressure of the one-way valve 62 between the reconstitution liquid tank 21 and the amplification reaction tank 31.

[0042] The first opening and closing member 33 includes a first paraffin wax 331 which is pre-fixed in the connecting groove 12 between the amplification reaction tank 31 and the diluent tank 41. The melting point of the first paraffin wax 331 is higher than the amplification reaction temperature. In the embodiment of the present application, the first paraffin wax 331 is a medium melting point paraffin wax, and semi-refined paraffin wax / full-refined paraffin wax, etc. can be used. The melting point of the first paraffin wax 331 is 55 degrees Celsius.

[0043] The second opening and closing member 43 includes a second paraffin wax 431 which is pre-fixed in the connecting groove 12 between the diluent tank 41 and the detection tank 51. The melting point of the second paraffin wax 431 is higher than that of the first paraffin wax 331. In the embodiment of the present application, the second paraffin wax 431 is a high melting point paraffin wax, and microcrystalline paraffin wax can be used. The melting point of the second paraffin wax 431 is 65 degrees Celsius.

[0044] In other embodiments, the first opening and closing member 33 and the second opening and closing member 43 can be plugging sliders. A sliding groove communicating with the connecting groove 12 is provided on the detection card body 1. The plugging slider can slide from the sliding groove into the connecting groove 12. When the plugging slider opens the connecting groove 12, the cover of the plugging slider covers the opening of the connecting groove 12 to prevent the sample liquid from overflowing.

[0045] In other embodiments, the second opening and closing member 43 can also be a one-way valve 62. The opening pressure of this one-way valve 62 is greater than the opening pressure of the one-way valve 62 between the re-melting liquid tank 21 and the amplification reaction tank 31. After the sample liquid dilution is completed, by continuously pressing the one-way pressing cap 61, the one-way valve 62 can be opened, and the sample liquid can be smoothly discharged into the detection tank 51.

[0046] After on-site sampling, the pine wood can be soaked to obtain pine wood nematode liquid or the pine wood chips can be directly scraped. The pine wood nematode liquid or the pine wood chips are added to a special reagent. The special reagent is a pine wood nematode lysis buffer and a pine wood nematode digestive enzyme, and the special reagent is heated to 95 °C and kept for 5 minutes to obtain a sample liquid;

[0047] The one-way pressing cap 61 is removed from the sample liquid injection port 11, and then the sample liquid is injected into the sample liquid injection port 11, and then the one-way pressing cap 61 is reinstalled. By squeezing the one-way pressing cap 61, the sample liquid can be injected into the re-melting liquid tank 21 to be re-melted with the re-melting liquid 22. After the re-melting is completed, the one-way pressing cap 61 is pressed again to increase the air pressure, and the sample liquid and the re-melting liquid 22 are injected into the amplification reaction tank 31. Then, the detection card body 1 is heated. The heating temperature is 40 degrees Celsius and kept for 20 minutes to melt the freeze-dried ball 32 and mix it with the sample liquid and the re-melting liquid 22 to complete the amplification reaction;

[0048] After the amplification reaction is completed, continue to heat the test card body 1 at a temperature of 55 degrees for 2 minutes to melt the first paraffin wax 331. Since the air pressure in the amplification reaction tank 31 is higher than that in the diluent tank 41, the sample solution can automatically flow into the diluent tank 41 for dilution. Moreover, the chemical inertness of the first paraffin wax 331 makes it not mix with the sample solution after melting, which can avoid interfering with the test results;

[0049] After the sample solution is diluted, continue to heat the test card body 1 at a temperature of 65 degrees for 2 minutes to melt the second paraffin wax 431. Since the air pressure in the diluent tank 41 is higher than that in the test tank 51, part of the diluted sample solution can automatically flow into the test tank 51. The test strip 52 absorbs the sample solution, and the tester can make a judgment according to the display of the test strip 52 to detect whether there are harmful pine wood nematodes in the sample solution. It should be noted that during the operation, if the sample solution cannot flow normally, the one-way pressing cap 61 can also be pressed continuously to inject gas to push the sample solution to flow.

[0050] It enables the sampling personnel to detect pine wood nematodes on-site during sampling and obtain accurate test results. Moreover, the operation is simple and convenient for detection. In addition, this disposable test card is convenient to carry, and there is no need for transfer during the detection process of the sample solution, reducing the situation of sample solution being contaminated. Similarly, the re-melting unit 2, the amplification reaction unit 3, the dilution unit 4, and the detection unit 5 are designed in a grooved manner to make the surface of the reagent card flat, reduce the occupied space, and be more convenient to carry.

[0051] The embodiment of the present application also discloses a portable pine wood nematode DNA detection module.

[0052] Such as Figure 2 and Figure 3, the portable pine wilt nematode DNA detection module includes the portable pine wilt nematode DNA detection card of the embodiment of the present application and a heating unit 7. The heating unit 7 includes a housing 71. Two slots 711 are formed in the housing 71, and both of the two slots 711 extend from one side of the housing 71. An operation port 712 corresponding to the slot 711 is formed in the housing 71. The operation port 712 penetrates to the top of the housing 71 and communicates with the slot 711. An HMI 74 is arranged on the housing 71, a control module 75 is arranged inside the housing 71, two heating sheets 72 are installed inside the housing 71, and the two heating sheets 72 are installed at the bottom of the slot 711. A power module 73 is also arranged inside the housing 71. The power module 73 is powered by an installed lithium battery. In other embodiments, the power module 73 can also be set as a built-in energy storage battery, and a USB socket is provided for charging. The control module 75 is connected to the heating sheet 72 and controls the heating sheet 72 to heat to different temperatures. The area of the slot 711 facing the heating sheet 72 is the heating area. The detection card body 1 can be inserted into the slot 711. When the detection card body 1 is inserted properly, the amplification reaction unit 3 and the dilution unit 4 are exactly located in the heating area.

[0053] It can heat the detection card body 1 at different temperatures. The power module 73 provides power without the need for an external power source, which is convenient for the detection personnel to carry and facilitates heating operations at the sampling site. The setting of multiple slots 711 and multiple heating sheets 72 can realize heating multiple detection card bodies 1 simultaneously, improve the detection efficiency, and the detection card body 1 can be fixed directly after being inserted into the slot 711. The sample liquid can be injected into the detection card body 1 through the operation port 712, which is convenient for operation.

[0054] The above are all the preferred embodiments of the present application. The protection scope of the present application is not limited hereby. Therefore, all equivalent changes made according to the structure, shape, and principle of the present application should be covered within the protection scope of the present application.

Claims

1. A portable pine wood nematode DNA detection card, characterized in that: The invention comprises a detection card body (1), on which a sample liquid injection port (11), a remelting unit (2), an amplification reaction unit (3), a dilution unit (4) and a detection unit (5) are arranged, wherein the sample liquid injection port (11) is connected to the remelting unit (2), the remelting unit (2) is connected to the amplification reaction unit (3), the amplification reaction unit (3) is connected to the dilution unit (4), and the dilution unit (4) is connected to the detection unit (5); and the detection card body (1) is provided with a control component (6) for controlling the flow of liquid from the sample liquid injection port (11) to the detection unit (5).

2. The portable pine wood nematode DNA detection card according to claim 1, characterized in that: The remelting unit (2) comprises a remelting liquid tank (21) provided on the detection card body (1), the remelting liquid tank (21) contains remelting liquid (22), and the slot of the remelting liquid tank (21) is sealed by a sealing film.

3. The portable pine wood nematode DNA detection card according to claim 1, characterized in that: The amplification reaction unit (3) comprises an amplification reaction groove (31) provided on the detection card body (1), wherein the amplification reaction groove (31) contains freeze-dried balls (32), the groove opening of the amplification reaction groove (31) is sealed by a sealing film, and a first opening and closing member (33) is provided at the discharge point of the amplification reaction groove (31).

4. The portable pine wood nematode DNA detection card according to claim 3, characterized in that: The first opening and closing member (33) includes a first paraffin wax (331), and the melting point of the first paraffin wax (331) is higher than the amplification reaction temperature.

5. The portable pine wood nematode DNA detection card according to claim 4, characterized in that: The dilution unit (4) comprises a dilution liquid groove (41) provided on the detection card body (1), wherein the dilution liquid groove (41) contains a dilution liquid (42), the groove opening of the dilution liquid groove (41) is sealed by a sealing film, and a second opening and closing member (43) is provided at the discharge point of the dilution liquid groove (41).

6. The portable pine wood nematode DNA detection card according to claim 5, characterized in that: The second opening and closing member (43) includes a second paraffin wax (431), and the melting point of the second paraffin wax (431) is higher than the melting point of the first paraffin wax (331).

7. The portable pine wood nematode DNA detection card according to claim 1, characterized in that: The detection unit (5) comprises a detection slot (51) provided on the detection card body (1), a test strip (52) being inserted into the detection slot (51), and a slot opening of the detection slot (51) being sealed by a sealing film.

8. The portable pine wood nematode DNA detection card according to claim 1, characterized in that: The control component (6) comprises a one-way pressing cap (61), which is detachably arranged on the sample liquid injection port (11), and a first one-way valve (62) is arranged at the connection between the sample liquid injection port (11) and the remelting unit (2), and at the connection between the remelting unit (2) and the amplification reaction unit (3).

9. A portable pine wood nematode DNA detection module, characterized in that: The portable pine wood nematode DNA detection card comprises any one of claims 4 to 6 and a heating unit (7), wherein the heating unit (7) comprises a casing (71), a heating plate (72) is arranged in the casing (71), a control module (75) for controlling the heating plate (72) to perform heating at different temperatures is arranged in the casing (71), a power module (73) is arranged in the casing (71), and the power module (73) is connected to the control module (75).

10. The portable pine wood nematode DNA detection module according to claim 9, characterized in that: At least two slots (711) are provided in the housing (71), the number of the heating plates (72) is the same as and corresponds to the number of the slots (711), the area directly opposite the slots (711) and the heating plates (72) is a heating zone, the detection card body (1) can be inserted into the slots (711), when the detection card body (1) is inserted, the amplification reaction unit (3) and the dilution unit (4) are located in the heating zone, and at least two operation ports (712) are provided on the housing (71), and the operation ports (712) correspond to and are connected with the slots (711) one by one.

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