CN106771193B - A kind of immunochromatographytest test kit of herpes simplex virus type II IgM antibody - Google Patents

Abstract

The invention belongs to the field of biomedicine, and in particular relates to an immunochromatographic detection kit for herpes simplex virus type II IgM antibody. The kit is assembled from a reagent strip and a plastic card. The reagent strip includes a bottom plate on which a reaction film with a detection line and a quality control line is pasted. The end of the reaction film close to the detection line is sequentially superimposed and pasted with a colloidal gold pad, The sample pad, the end of the reaction membrane near the quality control line is superimposed with an absorbent pad, the detection line is coated with herpes simplex virus type II antigen, the quality control line is coated with goat anti-mouse IgG antibody, and the colloidal gold pad is coated with colloidal gold markers mouse anti-human IgM monoclonal antibody. The kit of the invention can significantly reduce the influence of environmental factors (such as low temperature and high humidity) on the detection process and results, and ensure the sensitivity, accuracy and stability of the detection results.

Description

A kind of immunochromatographic detection kit of herpes simplex virus type II IgM antibody

technical field

The invention belongs to the field of biomedicine, and in particular relates to an immunochromatographic detection kit for herpes simplex virus type II IgM antibody.

Background technique

Herpes Simplex Virus (HSV) belongs to the human herpesvirus α subfamily, is a DNA double-stranded virus, and humans are its only natural host. According to the different antigenic properties, it can be divided into herpes simplex virus type I (HSV-1) type and herpes simplex virus type II (HSV-2 type), and the gene homologous sequence between the two is about 50%. The HSV-1 type usually infects the parts above the waist such as the mouth, face, and eyes, while the HSV-2 type usually infects the parts below the genitals and the perianal area. The typical clinical manifestations of herpes are clustered blisters and pustules on the skin and mucous membranes. Sores and scabs.

HSV-2 is the most common cause of genital herpes, which has become one of the most prevalent sexually transmitted infections. Pregnant women infected with herpes simplex virus can easily cause fetal malformation or herpes virus encephalitis in babies after delivery, which can cause serious disability. In recent years, the prevalence of genital herpes, especially recurrent genital herpes, has increased significantly in both developed and developing countries. More than 80% of HSV-2 infected persons do not show symptoms of genital herpes, or have no significant symptoms, and are not easy to be found, but they can still intermittently detoxify and infect others without being easy to detect. Both HSV-specific and non-specific antibodies appear within the first few weeks after infection. Therefore, detection of HSV-2-specific IgM antibodies plays an important role in silent infection and epidemiological investigations.

Currently, laboratory diagnostic methods for HSV include cell culture, antigen detection, PCR, and serum antibody detection. Cell culture and PCR methods have high requirements on experimental conditions such as experimental sites and personnel, and cannot be carried out in most laboratories. The direct immunofluorescence or ELISA methods for detecting antigens need to rely on corresponding instruments and equipment, and most of the reagents are imported and expensive. Serum antibody detection, such as the commonly used colloidal gold immunochromatography, is widely used due to its strong specificity, high sensitivity, and simple operation. However, the colloidal gold immunochromatography is greatly affected by environmental and human factors, such as When the temperature is low (such as winter), the chromatography speed is slow, and the reaction time needs to be extended, otherwise it is easy to cause missed detection; or it is placed in the air for a long time, resulting in moisture, or when the ambient humidity is too high, the capillary action on the membrane is strengthened, Dot film is easy to cause T line, C line to widen or even spread, affecting the detection process and results. Therefore, it is necessary to optimize the kit for detecting HSV-2 type-specific IgM antibody by colloidal gold immunochromatography to obtain a herpes simplex virus type II immunochromatographic detection reagent that is less affected by environmental factors and has high detection stability. box.

Contents of the invention

In order to solve the problems existing in the prior art (such as colloidal gold immunochromatography is greatly affected by environmental factors, resulting in decreased sensitivity and stability, etc.), the invention provides an immunochromatographic detection of herpes simplex virus type II IgM antibody The kit, which can significantly reduce the impact of environmental factors on the detection process and results, and ensure the sensitivity, accuracy and stability of the detection results.

The invention provides an immunochromatographic detection kit for herpes simplex virus type II IgM antibody, the kit is assembled from a reagent strip and a plastic card, the reagent strip includes a bottom plate, and the bottom plate is pasted with a The reaction membrane of the detection line (T) and the quality control line (C), the end of the reaction membrane close to the detection line (T) is superimposed and pasted with a colloidal gold pad and a sample pad in turn, and the reaction membrane is close to the quality control line (C) One end of the reaction membrane is superimposed and pasted with an absorbent pad, the detection line (T) on the reaction membrane is coated with herpes simplex virus type II antigen, the quality control line (C) on the reaction membrane is coated with goat anti-mouse IgG antibody, the The colloidal gold pad is coated with colloidal gold-labeled mouse anti-human IgM monoclonal antibody.

In the kit of the present invention, the reaction membrane is prepared by the following method: dilute the herpes simplex virus type II antigen with a buffer to a coating concentration of 0.8-1.2 mg/ml, and dilute the goat anti-mouse IgG antibody to a coating concentration of 0.8-1.2 mg/ml. The concentration is 1.5-1.8mg/ml, and the detection line of herpes simplex virus type II antigen and the quality control line of goat anti-mouse IgG antibody are respectively drawn on the reaction membrane. After marking, the reaction membrane is placed in a drying room at a temperature of 20-25°C , the humidity is less than 30%, dry for 4 to 5 hours;

The colloidal gold pad is prepared by the following method: a colloidal gold solution with a diameter of 20nm is prepared by the chloroauric acid-trisodium citrate reduction method, and after the preparation is completed, 100ml of the colloidal gold solution is placed in a beaker and mixed with 0.2MK ₂ CO ₃ Adjust to pH 8.0, add 2.0mg mouse anti-human IgM monoclonal antibody to 100ml colloidal gold solution, then slowly add 0.2M K ₂ CO ₃ to adjust the colloidal gold solution to pH 9.5, add K ₂ CO ₃ for 5 minutes , then stirred at room temperature for 30 minutes, added bovine serum albumin with a final concentration of 10mg/ml, 1mg/ml polyvinylpyrrolidone for blocking for 20-30 minutes, centrifuged at 12000r/m for 30 minutes, discarded the supernatant, and reconstituted to 50ml with buffer, Spread 1ml of the solution evenly on the glass fiber membrane at a ratio of 20cm ² , and then place it in a drying room at a temperature of 20-25°C and a humidity of less than 30%, and dry for 4-5 hours.

Further, the buffer solution is prepared by the following method: Weigh 0.06g of NaH ₂ PO ₄ ·2H ₂ O and 0.58g of Na ₂ HPO ₄ ·12H ₂ O, add 80.0 mL of purified water, stir to fully dissolve Add 0.80-0.90 g of NaCl and 0.5-2.0 g of alkyl glucoside, mix thoroughly, and set the volume to 100 ml, and measure the pH value to be 7.5-8.0.

Still further, the carbon chain length of the alkyl glycoside is C8-C10.

Further, the reaction membrane is a nitrocellulose membrane.

Further, the sample pad is a glass fiber membrane.

In the technical scheme of the present invention, alkyl glycoside is a kind of nonionic surfactant, which has the characteristics of both nonionic and anionic surfactants, and the inventor unexpectedly found that adding alkyl glycoside in phosphate buffer, And it is applied to the preparation of reaction membrane and colloidal gold pad at the same time, which can effectively improve the chromatography speed of the kit in low temperature environment, and weaken the influence of ambient humidity on the detection process and results, thereby improving the accuracy and stability of the detection results . This may be due to the non-ionic and anionic properties of alkyl glycosides. On the one hand, it can accelerate the speed of colloidal gold-labeled particles and ligands to form complexes during detection, as well as the capture speed of T-line and C-line, thereby increasing the chromatography speed. , so even in a low temperature environment, the detection reaction time is within 20 minutes; on the other hand, it can improve the stability of colloidal gold-labeled particles and complexes, maintain the stability of T-line and C-line, and greatly weaken the influence of environmental humidity. First, it can prolong its service life, and second, it can avoid the influence of the detection process and inaccurate detection results due to moisture, so as to ensure the sensitivity, accuracy and stability of the detection results.

The detection method of the kit of the present invention is: 1) balance the detection reagent and the sample to room temperature, take out the test paper box, and lay it flat; 2) 10 μl of serum and plasma samples, when the sample is whole blood, draw 20ul of the sample and add it to the sample hole , and immediately add 100 μL of sample diluent (normal saline or PBS) to the lower buffer well, and the result can be judged within 5 to 20 minutes. 3) Analysis of test results: Positive: Two bands of purple-red quality control line (C line) and test line (T-line) appear; Negative: only one band of purple-red quality control line (C line) appears; Invalid: no The appearance of purple-red quality control line (C line) strips indicates that the operation process is incorrect or the reagent has been deteriorated and damaged. The experiment is invalid, and the test result after 60 minutes is also invalid.

Therefore, compared with prior art, the advantage of the present invention is:

The immunochromatographic detection kit of herpes simplex virus type II IgM antibody of the present invention adds the component of alkyl glycoside by optimizing the buffer solution, and simultaneously coats herpes simplex virus type II antigen and goat anti-mouse IgG antibody Concentration, labeling concentration of mouse anti-human IgM monoclonal antibody, and pH value during colloidal gold labeling are adjusted and optimized to significantly reduce the impact of environmental factors on the detection process, speed up chromatography in low temperature environments, and improve the moisture resistance of the kit , so as to ensure the sensitivity, accuracy and stability of the test results, and prolong the service life of the kit.

Detailed ways

The present invention will be further described in detail below. It should be pointed out that the following description is only an illustration of the technical solutions claimed in the present invention, and is not any limitation to these technical solutions. The protection scope of the present invention shall be determined by the contents described in the appended claims.

In the examples and comparative examples of the present invention, the herpes simplex virus type II antigen was obtained by culturing and purifying the herpes simplex virus type II G strain, and other raw materials were obtained from the market, such as goat anti-mouse IgG antibody purchased from Hangzhou Qitai Biotechnology Co., Ltd. Co., Ltd., mouse anti-human IgM monoclonal antibody was purchased from Shanghai Dingjie Biotechnology Co., Ltd., and C8-C10 alkyl glycosides (active substance 52.0 wt%) were purchased from Guangzhou Xilu Chemical Co., Ltd.

Embodiment 1, the immunochromatographic detection kit of herpes simplex virus type II IgM antibody of the present invention

The test kit of the present embodiment is assembled by a reagent strip and a plastic card. The width of the reagent strip is 4 mm, and includes a base plate on which a test line (T) and a quality control line (C) are pasted on the base plate. A colloidal gold pad and a sample pad are superimposed and pasted on one end of the reaction film close to the detection line (T) in sequence, one end of the T line overlaps 1/5 of the colloidal gold pad, and the sample pad overlaps 1/5 of the colloidal gold pad. Paste at 3 places; the end of the reaction film close to the quality control line (C) is superimposed and pasted with an absorbent pad, and the 1/10 place of the overlapping suction pad is pasted, and the detection line (T) on the reaction film is coated with herpes simplex Virus type II antigen, the quality control line (C) on the reaction membrane is coated with goat anti-mouse IgG antibody, and the mouse anti-human IgM monoclonal antibody labeled with colloidal gold is coated on the colloidal gold pad.

The reaction membrane is prepared by the following method: the herpes simplex virus type II antigen is diluted to a coating concentration of 0.8 mg/ml with a buffer, and the goat anti-mouse IgG antibody is diluted to a coating concentration of 1.5 mg/ml. Scribe the detection line of herpes simplex virus type II antigen and the quality control line of goat anti-mouse IgG antibody on the membrane respectively. After scribing, place the reaction membrane in a drying room at a temperature of 20°C and a humidity of less than 30% for 5 hours;

The colloidal gold pad is prepared by the following method: a colloidal gold solution with a diameter of 20nm is prepared by the chloroauric acid-trisodium citrate reduction method, and after the preparation is completed, 100ml of the colloidal gold solution is placed in a beaker and mixed with 0.2MK ₂ CO ₃ Adjust to pH 8.0, add 2.0mg mouse anti-human IgM monoclonal antibody to 100ml colloidal gold solution, then slowly add 0.2M K ₂ CO ₃ to adjust the colloidal gold solution to pH 9.5, add K ₂ CO ₃ for 5 minutes , then stirred at room temperature for 30 minutes, added bovine serum albumin with a final concentration of 10mg/ml, blocked with 1mg/ml polyvinylpyrrolidone for 20 minutes, centrifuged at 12000r/m for 30 minutes, discarded the supernatant, reconstituted with buffer to 50ml, and pressed 1ml Spread the solution evenly on the glass fiber membrane at a ratio of 20 cm ² , and then place it in a drying room at a temperature of 20°C and a humidity of less than 30%, and dry for 5 hours.

The buffer solution is prepared by the following method: Weigh 0.06g of NaH ₂ PO ₄ ·2H ₂ O and 0.58g of Na ₂ HPO ₄ ·12H ₂ O, add 80.0mL of purified water, stir to fully dissolve, then add 0.80g NaCl and 0.5 g of alkyl glucoside, mix well, make the volume to 100 ml, and measure the pH value to be 7.5.

The carbon chain length of the alkyl glycoside is C8-C10; the reaction membrane is a nitrocellulose membrane; and the sample pad is a glass fiber membrane.

Embodiment 2, the immunochromatographic detection kit of herpes simplex virus type II IgM antibody of the present invention

The test kit of the present embodiment is assembled by a reagent strip and a plastic card. The width of the reagent strip is 4 mm, and includes a base plate on which a test line (T) and a quality control line (C) are pasted on the base plate. A colloidal gold pad and a sample pad are superimposed and pasted on one end of the reaction film close to the detection line (T) in sequence, one end of the T line overlaps 1/5 of the colloidal gold pad, and the sample pad overlaps 1/5 of the colloidal gold pad. Paste at 3 places; the end of the reaction film close to the quality control line (C) is superimposed and pasted with an absorbent pad, and the 1/10 place of the overlapping suction pad is pasted, and the detection line (T) on the reaction film is coated with herpes simplex Virus type II antigen, the quality control line (C) on the reaction membrane is coated with goat anti-mouse IgG antibody, and the mouse anti-human IgM monoclonal antibody labeled with colloidal gold is coated on the colloidal gold pad.

The reaction membrane is prepared by the following method: the herpes simplex virus type II antigen is diluted to a coating concentration of 1.2 mg/ml with a buffer, and the goat anti-mouse IgG antibody is diluted to a coating concentration of 1.8 mg/ml. Scribe the detection line of herpes simplex virus type II antigen and the quality control line of goat anti-mouse IgG antibody on the membrane respectively. After scribing, place the reaction membrane in a drying room at a temperature of 25°C and a humidity of less than 30% for 4 hours;

The colloidal gold pad is prepared by the following method: a colloidal gold solution with a diameter of 20nm is prepared by the chloroauric acid-trisodium citrate reduction method, and after the preparation is completed, 100ml of the colloidal gold solution is placed in a beaker and mixed with 0.2MK ₂ CO ₃ Adjust to pH 8.0, add 2.0mg mouse anti-human IgM monoclonal antibody to 100ml colloidal gold solution, then slowly add 0.2M K ₂ CO ₃ to adjust the colloidal gold solution to pH 9.5, add K ₂ CO ₃ for 5 minutes , then stirred at room temperature for 30 minutes, added bovine serum albumin with a final concentration of 10mg/ml, blocked with 1mg/ml polyvinylpyrrolidone for 30 minutes, centrifuged at 12000r/m for 30 minutes, discarded the supernatant, reconstituted with buffer to 50ml, and pressed 1ml Spread the solution evenly on the glass fiber membrane at a ratio of 20 cm ² , and then place it in a drying room at a temperature of 25°C and a humidity of less than 30%, and dry for 5 hours.

The buffer solution is prepared by the following method: Weigh 0.06g of NaH ₂ PO ₄ ·2H ₂ O and 0.58g of Na ₂ HPO ₄ ·12H ₂ O, add 80.0mL of purified water, stir to fully dissolve, then add 0.90g NaCl and 2.0 g of alkyl glucosides, mix well, make the volume to 100 ml, and measure the pH value to be 8.0.

The carbon chain length of the alkyl glycoside is C8-C10; the reaction membrane is a nitrocellulose membrane; and the sample pad is a glass fiber membrane.

Embodiment 3, the immunochromatographic detection kit of herpes simplex virus type II IgM antibody of the present invention

The test kit of the present embodiment is assembled by a reagent strip and a plastic card. The width of the reagent strip is 4 mm, and includes a base plate on which a test line (T) and a quality control line (C) are pasted on the base plate. A colloidal gold pad and a sample pad are superimposed and pasted on one end of the reaction film close to the detection line (T) in sequence, one end of the T line overlaps 1/5 of the colloidal gold pad, and the sample pad overlaps 1/5 of the colloidal gold pad. Paste at 3 places; the end of the reaction film close to the quality control line (C) is superimposed and pasted with an absorbent pad, and the 1/10 place of the overlapping suction pad is pasted, and the detection line (T) on the reaction film is coated with herpes simplex Virus type II antigen, the quality control line (C) on the reaction membrane is coated with goat anti-mouse IgG antibody, and the mouse anti-human IgM monoclonal antibody labeled with colloidal gold is coated on the colloidal gold pad.

The reaction membrane is prepared by the following method: the herpes simplex virus type II antigen is diluted to a coating concentration of 1.0 mg/ml with a buffer, and the goat anti-mouse IgG antibody is diluted to a coating concentration of 1.6 mg/ml. Scribe the detection line of herpes simplex virus type II antigen and the quality control line of goat anti-mouse IgG antibody on the membrane respectively. After scribing, place the reaction membrane in a drying room at a temperature of 20°C and a humidity of less than 30% for 5 hours;

The colloidal gold pad is prepared by the following method: a colloidal gold solution with a diameter of 20nm is prepared by the chloroauric acid-trisodium citrate reduction method, and after the preparation is completed, 100ml of the colloidal gold solution is placed in a beaker and mixed with 0.2MK ₂ CO ₃ Adjust to pH 8.0, add 2.0mg mouse anti-human IgM monoclonal antibody to 100ml colloidal gold solution, then slowly add 0.2M K ₂ CO ₃ to adjust the colloidal gold solution to pH 9.5, add K ₂ CO ₃ for 5 minutes , then stirred at room temperature for 30 minutes, added bovine serum albumin with a final concentration of 10mg/ml, blocked with 1mg/ml polyvinylpyrrolidone for 25 minutes, centrifuged at 12000r/m for 30 minutes, discarded the supernatant, reconstituted to 50ml with buffer, and pressed 1ml Spread the solution evenly on the glass fiber membrane at a ratio of 20 cm ² , and then place it in a drying room at a temperature of 20°C and a humidity of less than 30%, and dry for 5 hours.

The buffer solution is prepared by the following method: Weigh 0.06g of NaH ₂ PO ₄ ·2H ₂ O and 0.58g of Na ₂ HPO ₄ ·12H ₂ O, add 80.0mL of purified water, stir to fully dissolve, then add 0.8g NaCl and 1.0 g of alkyl glucoside, mix well, make the volume to 100 ml, and measure the pH value to be 7.8.

The carbon chain length of the alkyl glycoside is C8-C10; the reaction membrane is a nitrocellulose membrane; and the sample pad is a glass fiber membrane.

Comparative example 1

Compared with Example 3, the only difference in this comparative example is that no alkyl glycoside is added to the buffer.

Comparative example 2

Compared with Example 3, the only difference in this comparative example is that 4.0 g of alkyl glycosides are added to the buffer.

Test example one

Using the kits of Examples 1-3 and Comparative Examples 1-2, and a commercially available kit (herpes simplex virus type II (IgM) antibody detection kit, purchased from Fujian Mingxi Haitian Lanbo Biotechnology Co., Ltd.), Under different environmental conditions, 12 positive serum samples of herpes simplex virus type II IgM antibody were detected at the same time, and the results were as follows:

1) The test results of the 12 cases of serum samples under normal temperature conditions (temperature: 25 ± 2°C, humidity: 58 ± 2%) of the kits of Examples 1-3 and Comparative Examples 1-2, and commercially available kits were all the same. It is effective, and the T-lines of 12 cases of serum samples are all dark purple, which is basically the same color as the C-line, and all show positive; Among them, the test kits of Examples 1-3 can determine the results within 4-6 minutes, and the comparative example 1-2 kits and commercially available kits can determine the results within 10-20 minutes.

2) See Table 1 below: The test results of the kits in Examples 1-3 are all effective and positive for 12 serum samples under low temperature conditions (temperature: 8±2°C, humidity: 58±2%) , the results can be judged within 20 minutes, and the T-lines of 12 serum samples were all dark purple, which was basically the same color as the C-line. However, both the kit of Comparative Example 1 and the commercially available kit had missed detection under low temperature environment, and 3 cases of positive samples were detected as negative, and the chromatographic speed was slow, the color development time was prolonged, and the detection line was light in color. , the sensitivity and accuracy decreased; while the kit of Comparative Example 2 was invalid in 2 cases under low temperature environment, and the color development time was also prolonged, the detection line showed light color, and the sensitivity and accuracy decreased.

Table 1 Test results of each kit under low temperature conditions (temperature: 8±2°C, humidity: 58±2%)

(Note: The color development time and color depth in the table are all for the detection line, "+" means light purple red, "++" means deep purple red)

3) See Table 2 below: The test results of the kits in Examples 1 to 3 are all effective for 12 serum samples under high humidity conditions (temperature: 25±2°C, humidity: 90±2%), and all show If it is positive, the result can be judged within 10-15 minutes, and the T-lines of the 12 serum samples are all dark purple, which is basically the same color as the C-line. However, the kit of Comparative Example 1 was invalid in 4 cases under high humidity conditions, and the T-line color development in the valid samples was lighter; the kit of Comparative Example 2 and the commercially available kit also had many cases of invalidation under high humidity conditions. , and detected 2 cases of false negatives, compared with the embodiment, the detection sensitivity and accuracy were significantly reduced.

In addition, under high-humidity conditions, the T-line and C-line of the kits of Examples 1-3 of the present invention remained clear and did not fade or widen within 60 minutes after coloring; while the kits of Comparative Example 1 and the commercially available The T-line and C-line of the kit began to fade, widen, and diffuse after color development, and the T-line and C-line of the kit in Comparative Example 2 began to fade, widen, and diffuse about 5 minutes after color development. The phenomenon of diffusion shows that the kit of the present invention can maintain the stability of T-line and C-line within a certain period of time under high-humidity conditions, prolonging the result judgment time.

Table 2 Test results of each kit under high humidity conditions (temperature: 25±2°C, humidity: 90±2%)

(Note: each kit is tested after being placed in a high-humidity environment for 30 minutes. The color development time and color depth in the table are based on the detection line, "+" means light purple red, "++" means deep purple red)

The summary of the present invention only exemplifies some specific embodiments of the claims, wherein the technical features recorded in one or more technical solutions can be combined with any one or more technical solutions, and the technical solutions obtained by these combinations It is also within the scope of protection of the present application, just as these combined technical solutions have been specifically recorded in the disclosure content of the present invention.

Claims (3)

1. An immunochromatographic detection kit of herpes simplex virus type II IgM antibody, said kit is assembled by a reagent strip and a plastic card, said reagent strip comprises a base plate, and said base plate is pasted with a detection line And the reaction film of the quality control line, the end of the reaction film close to the detection line is superimposed and pasted with a colloidal gold pad and the sample pad in turn, and the end of the reaction film close to the quality control line is superimposed and pasted with an absorbent pad. The line is coated with herpes simplex virus type II antigen, the quality control line on the reaction membrane is coated with goat anti-mouse IgG antibody, and the colloidal gold pad is coated with colloidal gold-labeled mouse anti-human IgM monoclonal antibody. in: The reaction membrane is prepared by the following method: the herpes simplex virus type II antigen is diluted to a coating concentration of 0.8-1.2 mg/ml with a buffer, and the goat anti-mouse IgG antibody is diluted to a coating concentration of 1.5-1.8 mg/ml. ml, respectively streak the detection line of herpes simplex virus type II antigen and the quality control line of goat anti-mouse IgG antibody on the reaction membrane. After marking, place the reaction membrane in a drying room at a temperature of 20~25°C and a humidity of less than 30%. 4~5 hours; The colloidal gold pad is prepared by the following method: a colloidal gold solution with a diameter of 20nm is prepared by the chloroauric acid-trisodium citrate reduction method, and after the preparation is completed, 100ml of the colloidal gold solution is placed in a beaker and mixed with 0.2MK ₂ CO ₃ Adjust to pH 8.0, add 2.0mg mouse anti-human IgM monoclonal antibody to 100ml colloidal gold solution, then slowly add 0.2M K ₂ CO ₃ to adjust the colloidal gold solution to pH 9.5, add K ₂ CO ₃ for 5 minutes , then stirred at room temperature for 30 minutes, added bovine serum albumin with a final concentration of 10mg/ml, 1mg/ml polyvinylpyrrolidone to block for 20-30 minutes, centrifuged at 12000r/m for 30 minutes, discarded the supernatant, and reconstituted to 50ml with buffer, Spread 1ml of the solution evenly on the glass fiber membrane at a ratio of 20cm ² , and then place it in a drying room at a temperature of 20~25°C and a humidity of less than 30%, and dry for 4~5 hours; The buffer solution is prepared by the following method: weigh 0.06g of NaH ₂ PO ₄ 2H ₂ O and 0.58g of Na ₂ HPO ₄ 12H ₂ O, add 80.0 mL of purified water, stir to fully dissolve, then add 0.80~ 0.90g of NaCl and 0.5~2.0g of alkyl glucoside, mix thoroughly, set the volume to 100ml, and measure the pH value to be 7.5~8.0; The carbon chain length of the alkyl glycoside is C8-C10. 2. The immunochromatographic detection kit of herpes simplex virus type II IgM antibody as claimed in claim 1, wherein the reaction membrane is a nitrocellulose membrane. 3. The immunochromatographic detection kit of herpes simplex virus type II IgM antibody as claimed in claim 1, wherein the sample pad is a glass fiber membrane.