Detection kit for monkey pox antigen

Abstract

The invention discloses a monkey pox antigen detection kit, and belongs to the technical field of virus detection.The monkey pox antigen detection kit comprises a PVC base plate, a sample pad, a colloidal gold pad, a nitrocellulose membrane and a water absorption pad, the sample pad, the colloidal gold pad, the nitrocellulose membrane and the water absorption pad are sequentially arranged on the PVC base plate, and the colloidal gold pad is coated with a monkey pox labeled antibody-colloidal gold compound and a chicken IgY colloidal gold compound; the nitrocellulose membrane comprises a test line and a control line which are arranged in parallel and spaced from each other, the test line is coated with a monkey pox coating antibody, and the control line is coated with a goat anti-chicken IgY monoclonal antibody. The monkey pox antigen in the human throat swab and the skin exudate is detected by a double-antigen sandwich method, and the method has the characteristics of simplicity and convenience in operation, rapidness in result obtaining, high accuracy, comprehensive sample coverage and the like.

Description

Technical Field

[0001] The invention belongs to the technical field of virus detection, and specifically relates to a monkeypox antigen detection kit. Background Art

[0002] Monkeypox virus (MPXV) belongs to the Poxviridae family and is a double-stranded DNA virus with a genome length of about 197 kb. The virus can cause monkeypox in humans and animals (a zoonotic infectious disease). The monkeypox virus also infects rodents such as dormice, squirrels, and Gambian kangaroos. The monkeypox virus is divided into two evolutionary branches: (1) Branch I, formerly known as the Central African branch or the Congo Basin branch; (2) Branch II, formerly known as the West African branch, is further divided into branches IIa and IIb.

[0003] The monkeypox virus is transmitted through close contact between people, through droplets, and through contact with objects contaminated by the virus. It can also be transmitted vertically through the placenta. Sexual transmission cannot be ruled out. The monkeypox virus mainly invades the human body through mucous membranes and broken skin. It is mainly transmitted through direct contact with the diseased skin or mucous membranes of the case. It can also be transmitted through contact with virus-contaminated objects, long-term close inhalation of respiratory droplets from the case, contact with respiratory secretions, lesion exudates, blood and other body fluids of infected animals, or infection and transmission through bites and scratches of infected animals.

[0004] As the number of monkeypox virus and clinical patients continues to increase and the infected population continues to change, the demand for its diagnosis, screening and monitoring continues to increase, and the workload of testing continues to increase. How to make clinical monkeypox antigen detection technology easier to operate and the methods more mature, and how to improve the sensitivity and specificity of the test results are of great significance to clinical monkeypox antigen detection. Summary of the invention

[0005] The object of the present invention is to provide a monkeypox antigen detection kit to solve the problem of inaccurate detection results of monkeypox antigen detection kits in the prior art.

[0006] The purpose of the present invention can be achieved through the following technical solutions:

[0007] A monkeypox antigen detection kit comprises a PVC base plate and a sample pad, a colloidal gold pad, a nitrocellulose membrane and a water-absorbing pad which are sequentially arranged on the PVC base plate, wherein the colloidal gold pad is coated with a monkeypox marker antibody-colloidal gold complex and a chicken IgY colloidal gold complex; the nitrocellulose membrane comprises a test line (C line) and a control line (T line) which are arranged in parallel and spaced apart from each other, wherein the test line is coated with a monkeypox coated antibody, and the control line is coated with a goat anti-chicken IgY monoclonal antibody.

[0008] Furthermore, the preparation method of the colloidal gold pad comprises the following steps:

[0009] S1. preparing monkeypox labeled antibody-colloidal gold complex;

[0010] S2, preparing chicken IgY colloidal gold complex;

[0011] S3. The monkeypox labeled antibody-colloidal gold complex and the chicken IgY colloidal gold complex are mixed uniformly at a volume ratio of 1:1 to obtain a mixed solution. The glass cellulose membrane is soaked in the mixed solution for 5 minutes, then taken out and dried at 37-45° C. for 16-24 hours to obtain a colloidal gold pad.

[0012] Furthermore, the method for preparing the monkeypox marker antibody-colloidal gold complex comprises the following steps:

[0013] Add K2CO3 solution to the colloidal gold solution, mix well, then add monkeypox labeled antibody, mix well, let stand for 30 minutes, add BSA (bovine serum albumin) solution to a final concentration of 1%, let stand for 30 minutes, centrifuge at 4°C, 10000rpm for 15 minutes, discard the supernatant to obtain the precipitate, add gold label diluent to resuspend, and obtain monkeypox labeled antibody-colloidal gold complex.

[0014] Furthermore, the usage ratio of colloidal gold solution, K2CO3 solution and monkeypox labeled antibody is 1mL:58-60uL:18uL, and the concentration of K2CO3 solution is 0.05M.

[0015] Furthermore, the monkeypox marker antibody is human anti-MPXV A29LmAb (39).

[0016] Furthermore, the mass fraction of the BSA (bovine serum albumin) solution is 10%.

[0017] Furthermore, the method for preparing the chicken IgY colloidal gold complex comprises the following steps:

[0018] Add K2CO3 solution to the colloidal gold solution, mix well, add chicken IgY antibody, mix well and let stand for 30 minutes, add blocking solution with a final concentration of 10%, let stand for 30 minutes, centrifuge at 4°C, 10000rpm for 15 minutes, discard the supernatant to obtain the precipitate, add gold label diluent to resuspend, and obtain chicken IgY colloidal gold complex.

[0019] Furthermore, the usage ratio of the colloidal gold solution, K2CO3 solution and chicken IgY antibody is 1mL:12uL:10ug, and the concentration of the K2CO3 solution is 0.2M.

[0020] Furthermore, the blocking solution is a 10% BSA (bovine serum albumin) solution.

[0021] Furthermore, the colloidal gold solution is a colloidal gold solution prepared by a citric acid reduction method and having a particle size of 530 nm and an OD value of 4.0 to 5.0.

[0022] Furthermore, the pH value of the gold label diluent was 8.4±0.2, and it included 20 mM tris, 0.3% PVPK-30, 1.0% Triton-X100, 0.2% Casein-Na, 5.0% trehalose, and 1.0% BSA.

[0023] Furthermore, the method for preparing the sample pad comprises the following steps:

[0024] Pour the sample pad treatment liquid into a container, add glass fiber 8965, tap it continuously to remove bubbles, then take it out and spin it to drain until no water drops, place it flat on a drying rack, and dry it at 37-45℃ for 16-24h to obtain a sample pad.

[0025] Furthermore, the pH value of the sample pad treatment solution is 8.4±0.2, and includes 10 mM PBS (phosphate buffer), 0.1% Casein-Na, 0.1% BSA, 0.5% Triton-X100, and 0.4% PVPK-30.

[0026] Furthermore, the method for preparing the nitrocellulose membrane comprises the following steps:

[0027] The monkeypox coated antibody was diluted to a concentration of 1 mg / mL with a coating buffer, and then sprayed on the test area of ​​the nitrocellulose membrane by a film sprayer at a spraying amount of 0.9-1.1 uL / cm, and then dried at 37-45°C for 16 hours to obtain a test line; the sheep anti-chicken IgY monoclonal antibody was diluted to 1.0 mg / mL with a 10mM PBS solution, and then sprayed on the control area of ​​the nitrocellulose membrane by a film sprayer at a spraying amount of 1.2 uL / cm, and then dried at 37-45°C for 16 hours to obtain a control line, thereby obtaining a nitrocellulose membrane.

[0028] Furthermore, the monkeypox coated antibody is human anti-MPXV A29LmAb (B4).

[0029] Furthermore, the pH value of the coating buffer is 7.8±0.2, and includes 20 mM tris, 100 mM NaCl, and 3% trehalose.

[0030] The preparation method of the above-mentioned monkeypox antigen detection kit comprises the following steps:

[0031] Turn on the dehumidifier, and let the humidity of the environment drop below 35%, then take out the required materials and assemble them as required. Tear off the sticker on the PVC base plate at the absorbent pad position, and stick the absorbent pad of the specified size, with one end aligned with the PVC base plate; tear off the stickers at the sample pad and colloidal gold pad positions, first cover one end of the colloidal gold pad on the nitrocellulose membrane by 1-2mm, then align one end of the sample pad with the rubber plate, and the other end of the sample pad covers the colloidal gold pad by 1-2mm, and the nitrocellulose membrane is pasted on the PVC base plate.

[0032] Furthermore, the method for using the above-mentioned monkeypox antigen detection kit comprises the following steps:

[0033] (1) Collection of throat swabs or skin exudate to obtain sampling swabs;

[0034] (2) Place the monkeypox antigen detection kit on a clean and horizontal surface, add 100uL of sample diluent into the extraction tube, touch one end of the sampling swab to the sample pad, rotate the sampling swab while rinsing it with sample diluent, then start the timer and read the result after 15 to 20 minutes. Results displayed after 30 minutes or more have no clinical significance.

[0035] Further, the diluent includes 0.7089% Na2HPO4·12H2O, 0.8689% NaCl, 0.4% Tween, 0.5% PVP-40, and 0.1% PC-300.

[0036] Further, the results are interpreted as follows:

[0037] Positive: Two red bands appear, one within the control line (T line) and the other within the test line (C line).

[0038] Negative: Only a red stripe appears on the control test line (C line).

[0039] Invalid: The control line test line (C line) cannot be displayed. Regardless of whether the test line control line (T line) has a red line, the test equipment is invalid.

[0040] Quality Control: The red line appearing in the test line (Line C) is the internal process control; it confirms sufficient sample size.

[0041] Furthermore, throat swabs were collected as follows:

[0042] Insert the sampling swab into the oral cavity to the pharynx, rotate the swab for 15-20 seconds and then remove it.

[0043] Furthermore, skin exudate was collected as follows:

[0044] Place the sampling swab on the skin exudate, rotate the swab for 15-20 seconds and then remove it.

[0045] Beneficial effects of the present invention:

[0046] The monkeypox antigen detection kit of the present invention adopts a highly specific antigen-antibody reaction principle and immunochromatographic analysis technology to detect monkeypox antigens in human throat swabs and skin exudates by a double antigen sandwich method, which has the following advantages:

[0047] 1. Easy to operate: The test strips of the present invention are user-friendly and can be tested without the need for specialized equipment or technicians.

[0048] 2. Rapid results: Test results can be obtained in about 15-20 minutes and can be read in 10 minutes, which helps to quickly understand the infection status.

[0049] 3. High accuracy: Clinical trials have shown that the detection method has a 100% positive and negative consistency rate, making it suitable for auxiliary testing in hospitals and physical examination institutions.

[0050] 4. Comprehensive sample coverage: Suitable for testing of throat swabs or skin exudates. Clinically verified sample equivalence is good, and the test results of different samples are consistent, making it suitable for testing in different situations. BRIEF DESCRIPTION OF THE DRAWINGS

[0051] The present invention will be further described below in conjunction with the accompanying drawings.

[0052] Figure 1 It is a schematic structural diagram of the monkeypox antigen detection kit of the present invention;

[0053] Figure 2 This is a diagram showing the positive test results of the monkeypox antigen detection kit of the present invention on a throat swab sample;

[0054] Figure 3 This is a diagram showing the positive test results of the monkeypox antigen detection kit of the present invention on skin exudate. DETAILED DESCRIPTION

[0055] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0056] Example 1

[0057] A monkeypox antigen detection kit, such as Figure 1As shown, it includes a PVC base plate 1 and a sample pad 2, a colloidal gold pad 3, a nitrocellulose membrane 4 and a water-absorbing pad 5 which are sequentially arranged on the PVC base plate 1, the colloidal gold pad 3 is coated with a monkeypox labeled antibody-colloidal gold complex and a chicken IgY colloidal gold complex; the nitrocellulose membrane includes a test line (C line) 6 and a control line (T line) 7 which are arranged in parallel and spaced apart from each other, the test line 6 is coated with a monkeypox coated antibody, and the control line 7 is coated with a sheep anti-chicken IgY monoclonal antibody.

[0058] The preparation method of the colloidal gold pad 3 comprises the following steps:

[0059] S1. preparing monkeypox labeled antibody-colloidal gold complex;

[0060] S2, preparing chicken IgY colloidal gold complex;

[0061] S3. The monkeypox labeled antibody-colloidal gold complex and the chicken IgY colloidal gold complex are mixed uniformly at a volume ratio of 1:1 to obtain a mixed solution. The glass cellulose membrane is soaked in the mixed solution for 5 minutes, then taken out and dried at 37° C. for 24 hours to obtain a colloidal gold pad 3.

[0062] The method for preparing the monkeypox labeled antibody-colloidal gold complex comprises the following steps:

[0063] To 1 mL of colloidal gold solution, add 58 uL of 0.05 M K2CO3 solution, mix well, add 18 uL of human anti-MPXV A29L mAb (39), mix well, let stand for 30 min, add 10% BSA (bovine serum albumin) solution to a final concentration of 1%, let stand for 30 min, centrifuge at 4°C, 10000 rpm for 15 min, discard the supernatant to obtain the precipitate, add the gold label diluent to resuspend, and obtain the monkeypox labeled antibody-colloidal gold complex.

[0064] The method for preparing the chicken IgY colloidal gold complex comprises the following steps:

[0065] Add 12uL of 0.2M K2CO3 solution to 1mL of colloidal gold solution, mix well, add 10ug chicken IgY antibody, mix well and let stand for 30 minutes, add blocking solution with a final concentration of 10%, let stand for 30 minutes, centrifuge at 4°C, 10000rpm for 15 minutes, discard the supernatant to obtain the precipitate, add gold label diluent to resuspend, and obtain chicken IgY colloidal gold complex.

[0066] The blocking solution is 10% BSA (bovine serum albumin) solution.

[0067] The colloidal gold solution is a colloidal gold solution prepared by a citric acid reduction method, with a particle size of 530 nm and an OD value of 4.0 to 5.0.

[0068] The pH value of the gold standard diluent was 8.4 ± 0.2, including 20 mM tris, 0.3% PVPK-30, 1.0% Triton-X100, 0.2% Casein-Na, 5.0% trehalose, and 1.0% BSA.

[0069] The method for preparing the sample pad 2 comprises the following steps:

[0070] Pour the sample pad treatment liquid into a container, add glass fiber 8965, tap it continuously to remove bubbles, then take it out and spin it to drain until no water drops, place it flat on a drying rack, and dry it at 37°C for 24 hours to obtain sample pad 2.

[0071] The pH value of the sample pad treatment solution was 8.4±0.2, and it included 10 mM PBS (phosphate buffer), 0.1% Casein-Na, 0.1% BSA, 0.5% Triton-X100, and 0.4% PVPK-30.

[0072] The method for preparing the nitrocellulose membrane 4 comprises the following steps:

[0073] The human anti-MPXV A29LmAb (B4) was diluted with coating buffer to a concentration of 1 mg / mL, and then sprayed on the test area of ​​the nitrocellulose membrane 4 by a film sprayer, with a spraying amount of 1.1 uL / cm, and then dried at 45°C for 16 hours to obtain a test line 6; the sheep anti-chicken IgY monoclonal antibody was diluted with 10mM PBS solution to 1.0 mg / mL, and then sprayed on the control area of ​​the nitrocellulose membrane 4 by a film sprayer, with a spraying amount of 1.2 uL / cm, and then dried at 37°C for 16 hours to obtain a control line 7, thereby obtaining a nitrocellulose membrane 4.

[0074] The coating buffer has a pH value of 7.8±0.2 and includes 20 mM tris, 100 mM NaCl, and 3% trehalose.

[0075] The preparation method of the above-mentioned monkeypox antigen detection kit comprises the following steps:

[0076] Turn on the dehumidifier, and after the humidity of the environment drops below 35%, take out the required materials and assemble them as required. Tear off the sticker on the PVC base plate 1 at the position of the absorbent pad 5, and paste the absorbent pad 5 of the specified size, with one end aligned with the PVC base plate 1; tear off the stickers at the positions of the sample pad 2 and the colloidal gold pad 3, first cover one end of the colloidal gold pad 3 on the nitrocellulose membrane 4 by 2mm, then align one end of the sample pad 2 with the rubber plate, and cover the other end of the sample pad 2 on the colloidal gold pad 3 by 2mm, and paste the nitrocellulose membrane 4 on the PVC base plate 1.

[0077] Example 2

[0078] A monkeypox antigen detection kit, compared with Example 1, the only difference is that the preparation method of the monkeypox marker antibody-colloidal gold complex is different. The preparation method of the monkeypox marker antibody-colloidal gold complex in this embodiment comprises the following steps:

[0079] To 1 mL of colloidal gold solution, add 60 uL of 0.05 M K2CO3 solution, mix well, then add 18 uL of human anti-MPXV A29L mAb (39), mix well, let stand for 30 min, add 10% BSA (bovine serum albumin) solution to a final concentration of 1%, let stand for 30 min, centrifuge at 4°C, 10000 rpm for 15 min, discard the supernatant to obtain the precipitate, add the gold label diluent to resuspend, and obtain the monkeypox labeled antibody-colloidal gold complex.

[0080] The mass fraction of the BSA (bovine serum albumin) solution is 10%.

[0081] The composition of the colloidal gold solution and the gold label diluent resuspension is the same as that in Example 1.

[0082] Example 3

[0083] A monkeypox antigen detection kit, compared with Example 1, the only difference is that the preparation method of the nitrocellulose membrane 4 is different. The preparation method of the nitrocellulose membrane 4 in this embodiment comprises the following steps:

[0084] The human anti-MPXV A29LmAb (B4) was diluted with coating buffer to a concentration of 1 mg / mL, and then sprayed on the test area of ​​the nitrocellulose membrane 4 by a film sprayer at a spraying amount of 0.9 uL / cm, and then dried at 45°C for 16 hours to obtain a test line 6; the sheep anti-chicken IgY monoclonal antibody was diluted with 10mM PBS solution to 1.0 mg / mL, and then sprayed on the control area of ​​the nitrocellulose membrane 4 by a film sprayer at a spraying amount of 1.2 uL / cm, and then dried at 37°C for 16 hours to obtain a control line 7, thereby obtaining a nitrocellulose membrane 4.

[0085] The coating buffer has a pH value of 7.8±0.2 and includes 20 mM tris, 100 mM NaCl, and 3% trehalose.

[0086] The monkeypox antigen detection kit obtained in Example 1 was tested for accuracy, and the specific process was as follows:

[0087] (1) Collection of throat swabs or skin exudate to obtain sampling swabs;

[0088] Throat swabs were collected as follows:

[0089] Insert the sampling swab into the oral cavity to the pharynx, rotate the swab for 15 seconds and then remove it.

[0090] Skin exudate is collected as follows:

[0091] Place the sampling swab on the skin exudate, rotate the swab for 20 seconds and then remove it.

[0092] (2) Place the monkeypox antigen detection kit obtained in Example 1 on a clean and horizontal plane, add 100 uL of sample diluent to the extraction tube, contact one end of the sampling swab with the sample pad 2, rotate the sampling swab while rinsing it with the sample diluent, and then start timing. Read the results after 15 to 20 minutes. Results displayed after 30 minutes or more have no clinical significance.

[0093] The diluent includes 0.7089% Na2HPO4·12H2O, 0.8689% NaCl, 0.4% Tween, 0.5% PVP-40, and 0.1% PC-300.

[0094] The results are interpreted as follows:

[0095] Positive: Two red bands appear, one within the control line 7 (T line) and the other within the test line 6 (C line).

[0096] Negative: Only a red stripe appears on the test line 6 (line C).

[0097] Invalid: Control line test line 6 (C line) cannot be displayed. Regardless of whether the test line control line 7 (T line) has a red line, the test equipment is invalid.

[0098] Test results such as Figure 2 and Figure 3 As shown, Figure 2 2301x-x indicates that the sample type is a throat swab. Figure 3 2307x-x means the sample type is skin exudate, and "+" means the test is positive. The more "+" there are, the stronger the positive result is. Figure 2 and Figure 3 It can be seen that the test kit of Example 1 was used to test 5 throat swabs and 5 skin exudates of monkeypox virus infected persons, and the test results were all positive, with a detection rate of 100%, indicating that the test kit has the characteristics of high accuracy and comprehensive sample coverage.

[0099] It should be noted that, in this article, relational terms such as first and second, etc. are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Moreover, the terms "include", "comprise" or any other variants thereof are intended to cover non-exclusive inclusion, so that a process, method, article or device including a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements inherent to such process, method, article or device.

[0100] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A monkeypox antigen detection kit, characterized in that: The invention comprises a PVC bottom plate and a sample pad, a colloidal gold pad, a nitrocellulose membrane and a water-absorbing pad which are arranged on the PVC bottom plate in sequence, wherein the colloidal gold pad is coated with a monkeypox marker antibody-colloidal gold complex and a chicken IgY colloidal gold complex; the nitrocellulose membrane comprises a test line and a control line which are arranged in parallel and spaced apart from each other, wherein the test line is coated with a monkeypox coated antibody, and the control line is coated with a sheep anti-chicken IgY monoclonal antibody.

2. A monkeypox antigen detection kit according to claim 1, characterized in that: The preparation method of the colloidal gold pad comprises the following steps: S1. preparing monkeypox labeled antibody-colloidal gold complex; S2, preparing chicken IgY colloidal gold complex; S3. The monkeypox labeled antibody-colloidal gold complex and the chicken IgY colloidal gold complex are mixed uniformly at a volume ratio of 1:1 to obtain a mixed solution. The glass cellulose membrane is soaked in the mixed solution for 5 minutes, then taken out and dried at 37-45° C. for 16-24 hours to obtain a colloidal gold pad.

3. A monkeypox antigen detection kit according to claim 2, characterized in that: The method for preparing the monkeypox labeled antibody-colloidal gold complex comprises the following steps: Add K2CO3 solution to the colloidal gold solution, mix well, then add monkeypox labeled antibody, mix well, let stand for 30 minutes, add BSA solution to a final concentration of 1%, let stand for 30 minutes, centrifuge at 4°C, 10000rpm for 15 minutes, discard the supernatant to obtain the precipitate, add gold label diluent to resuspend, and obtain monkeypox labeled antibody-colloidal gold complex.

4. A monkeypox antigen detection kit according to claim 3, characterized in that: The usage ratio of colloidal gold solution, K2CO3 solution and monkeypox labeled antibody is 1mL:58~60uL:18uL, and the concentration of K2CO3 solution is 0.05M.

5. A monkeypox antigen detection kit according to claim 3, characterized in that: The monkeypox marker antibody is human anti-MPXV A29L mAb (39).

6. A monkeypox antigen detection kit according to claim 2, characterized in that: The method for preparing the chicken IgY colloidal gold complex comprises the following steps: Add K2CO3 solution to the colloidal gold solution, mix well, add chicken IgY antibody, mix well and let stand for 30 minutes, add blocking solution with a final concentration of 10%, let stand for 30 minutes, centrifuge at 4°C, 10000rpm for 15 minutes, discard the supernatant to obtain the precipitate, add gold label diluent to resuspend, and obtain chicken IgY colloidal gold complex.

7. A monkeypox antigen detection kit according to claim 6, characterized in that: The usage ratio of the colloidal gold solution, K2CO3 solution and chicken IgY antibody is 1mL:12uL:10ug, and the concentration of the K2CO3 solution is 0.2M.

8. A monkeypox antigen detection kit according to claim 1, characterized in that: The method for preparing the nitrocellulose membrane comprises the following steps: The monkeypox coating antibody was diluted to a concentration of 1 mg / mL with a coating buffer, and then sprayed on the test area of ​​the nitrocellulose membrane by a film sprayer, and then dried at 37-45°C for 16 hours to obtain a test line; the sheep anti-chicken IgY monoclonal antibody was diluted to 1.0 mg / mL with a 10mM PBS solution, and then sprayed on the control area of ​​the nitrocellulose membrane by a film sprayer, and then dried at 37-45°C for 16 hours to obtain a control line, thereby obtaining a nitrocellulose membrane.

9. A monkeypox antigen detection kit according to claim 8, characterized in that: The monkeypox coated antibody is human anti-MPXV A29L mAb (B4).

10. A monkeypox antigen detection kit according to claim 1, characterized in that: The method for preparing the sample pad comprises the following steps: Pour the sample pad treatment liquid into a container, add glass fiber 8965, tap it continuously to remove bubbles, then take it out and spin it to drain until no water drops, place it flat on a drying rack, and dry it at 37-45℃ for 16-24h to obtain a sample pad.

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