Helicobacter pylori antibody typing fluorescence immunochromatography detection reagent card and application thereof

Abstract

The invention belongs to the field of fluorescence immunoassay, and particularly relates to a helicobacter pylori antibody typing immunochromatography detection reagent card and application thereof. The reagent card comprises a coating plate, a sample pad, a combination pad, a nitrocellulose membrane and absorbent paper, wherein the sample pad, the combination pad, the nitrocellulose membrane and the absorbent paper are sequentially overlapped and adhered to the coating plate. According to the reagent card, helicobacter pylori vacuolar cytotoxin (VacA) antibodies, cytotoxin-related protein (CagA) antibodies and urease antibodies in human serum, plasma whole blood or fingertip blood samples are qualitatively detected through an indirect method and a fluorescence immunochromatography method, the Urease antibodies comprise UreA and UreB, relative fluorescence signal intensities of the various antibodies are obtained through detection of an immunoassay instrument, and the relative fluorescence signal intensities of the various antibodies are compared with the relative fluorescence signal intensities of the various antibodies. And judging whether the helicobacter pylori is positive or negative according to the reference value, and distinguishing the helicobacter pylori type I from the helicobacter pylori type II. Meanwhile, the reagent card also improves the sensitivity and repeatability of the product, and can meet the detection of serum / plasma / whole blood / fingertip blood.

Description

Technical Field

[0001] The present application belongs to the field of fluorescent immunoassay, and specifically relates to a Helicobacter pylori antibody typing fluorescent immunochromatographic detection reagent card and its application. Background Art

[0002] Helicobacter pylori (H.pylori, HP) infection is the most common Gram-negative spiral bacteria in the world. More than half of the world's population is infected with Helicobacter pylori. Its prevalence is related to socioeconomic conditions. About 50% of the adult population in developed countries and about 90% of the population in developing countries are infected with this bacterium. Helicobacter pylori is an important pathogenic factor of chronic gastritis and peptic ulcer, and is closely related to the occurrence of gastric cancer. In 1994, the International Agency for Research on Cancer classified Helicobacter pylori infection as a Class I carcinogen. Helicobacter pylori belongs to the subfamily of Proteus, Campylobacterales, Helicobacteraceae, with a total of 20 species. This microorganism has the characteristics of microaerobiosis and is positive for catalase, oxidase and urease in most cases. Urease is a nickel-ion-dependent enzyme that catalyzes the hydrolysis of urea into ammonia and carbonic acid. It is the most abundant protein in Helicobacter pylori, accounting for 5-10% of the total HP protein. Urease helps the bacteria settle in the stomach by neutralizing gastric acid and provides ammonia for bacterial protein synthesis, playing an important role in the parasitism and pathogenicity of HP in the stomach. The molecular weight of urease is about 550KD, and its monomer is composed of two subunits, A and B, with molecular weights of about 30KD and 66KD respectively; among them, urease B subunit is the active subunit of urease, which is well conserved in various HP strains and is recognized as an ideal antigen for HP vaccine; HP vaccine targeting urease A subunit can also achieve good protection, so urease A and B subunits are both ideal antigens for HP antibody preparation.

[0003] Helicobacter pylori mainly contains two types of cytotoxins: one is the cytotoxin-associated protein CagA, and the other is the vacuolar cytotoxin VacA. Clinically, Helicobacter pylori infection is divided into two types: Type I is the cytotoxin-producing Helicobacter pylori strain, CagA(+) and VacA(+) strains, with a detection rate of 50% to 60%. This type of infection can cause more serious gastric mucosal lesions and is a predisposing factor for gastric cancer; Type II is Helicobacter pylori that does not produce cytotoxins, namely CagA(-) and VacA(-) strains. This type of Helicobacter pylori strain has low toxicity and generally only causes chronic superficial gastritis without clinical symptoms after infection. The virulence of Helicobacter pylori is classified by toxin detection, and Urease positivity further confirms Helicobacter pylori infection.

[0004] At present, the reagent products for typing detection include quantum dot immunochromatography and immunoblotting. The detection of quantum dot immunochromatography is relatively simple, but its sensitivity is still not enough for the detection of whole blood and fingertip blood; the immunoblotting method takes a long time to operate and has high requirements for technicians. Although the two methods provide convenience for Helicobacter pylori typing detection, their sensitivity is still not enough for the detection of whole blood and fingertip blood. Summary of the invention

[0005] In response to the above-mentioned problems, the present application provides a Helicobacter pylori typing fluorescent immunochromatographic detection reagent card and its application. The detection reagent card uses an indirect method and fluorescent immunochromatographic method to qualitatively detect Helicobacter pylori vacuolar cytotoxin (VacA) antibodies, cytotoxin-associated protein (CagA) antibodies and urease (Urease) antibodies in human serum, plasma, whole blood or fingertip blood samples.

[0006] On the one hand, the present application provides a Helicobacter pylori typing fluorescent immunochromatographic detection reagent card, comprising a coating plate, a sample pad, a conjugation pad, a nitrocellulose membrane and absorbent paper, wherein the sample pad, the conjugation pad, the nitrocellulose membrane and the absorbent paper are overlapped in sequence and pasted on the coating plate; a first detection line sprayed with VacA antigen, a second detection line sprayed with CagA antigen, a third detection line sprayed with a mixture of Ure A and Ure B antigens and a quality control line sprayed with sheep anti-chicken IgY antibody are arranged in parallel from one end of the conjugation pad of the nitrocellulose membrane to one end of the absorbent paper, and the first detection line, the second detection line, the third detection line and the quality control line are parallel to the overlap; a mixture of mouse anti-human IgG antibody and chicken IgY antibody labeled with fluorescent microspheres is sprayed on the conjugation pad.

[0007] Using the above technical solution, the test strip contains Helicobacter pylori vacuolar cytotoxin antigen, cytotoxin-associated protein antigen, urease A / B antigen and sheep anti-chicken IgY antibody in the quality control area (C) pre-fixed on the nitrocellulose membrane, and the marking pad contains pre-coated fluorescently labeled mouse anti-human IgG and chicken IgY antibodies.

[0008] When testing the sample, the Helicobacter pylori vacuolar cytotoxin antibody / cytotoxin-associated protein antibody / urease antibody in the sample first binds to the fluorescently labeled mouse anti-human IgG to form an immune complex. Under the action of immunochromatography, the complex and the sample flow inside the nitrocellulose membrane toward the absorbent paper. When the complex passes through the detection area (T), it binds to the coated Helicobacter pylori vacuolar cytotoxin antigen / cytotoxin-associated protein antigen / urease antigen, forming "Helicobacter pylori vacuolar cytotoxin antigen-Helicobacter pylori vacuolar cytotoxin antibody-fluorescently labeled mouse anti-human IgG", "cytotoxin-associated protein antigen-cytotoxin-associated protein antibody-fluorescently labeled mouse anti-human IgG", "urease antigen-urease antibody-fluorescently labeled mouse anti-human IgG" complexes and agglutinates. When passing through the quality control area (C), the fluorescently labeled chicken IgY antibody binds to the coated goat anti-chicken IgY antibody, forming a "goat anti-chicken IgY antibody-fluorescently labeled chicken IgY antibody" complex and agglutinates. The fluorescent particles emit fluorescent signals under the stimulation of excitation light. The concentrations of Helicobacter pylori vacuoling cytotoxin antibodies, cytotoxin-associated protein antibodies and urease antibodies in the sample show a certain correlation with the fluorescence signal intensity. The relative fluorescence signal intensity is detected by the immunoassay analyzer, and the positive and negative of Helicobacter pylori vacuoling cytotoxin antibodies, cytotoxin-associated protein antibodies and urease antibodies are judged according to the corresponding reference values ​​of Helicobacter pylori vacuoling cytotoxin antibodies, cytotoxin-associated protein antibodies and urease antibodies.

[0009] In certain embodiments, the amino acid sequence of the VacA antigen is shown as SEQ ID NO.1, the amino acid sequence of the CagA antigen is shown as SEQ ID NO.2, the amino acid sequence of the Ure A antigen is shown as SEQ ID NO.3, and the amino acid sequence of the Ure B antigen is shown as SEQ ID NO.4.

[0010] In certain embodiments, the concentration of the VacA antigen, CagA antigen, Ure A antigen, and Ure B antigen is 0.05-0.5 mg / mL, preferably 0.1-0.3 mg / mL; the concentration of the goat anti-chicken IgY antibody is 0.1-1 mg / mL, preferably 0.3-0.8 mg / mL.

[0011] Using the above technical solution, the present application screened urease A protein, urease B protein, vacuolating toxin protein and cytotoxin protein for pairing with mouse anti-human IgG, and optimized the sequence and the concentration of the sprayed antigen on the reagent card, effectively improving the sensitivity and repeatability of the detection.

[0012] In certain embodiments, the preparation method of the fluorescent microsphere-labeled mouse anti-human IgG antibody and chicken IgY antibody mixture is to label the mouse anti-human IgG antibody and chicken IgY antibody with fluorescent microspheres respectively, then dilute them with microsphere diluent, and mix them at a volume ratio of 1:20 to obtain a fluorescent microsphere-labeled mouse anti-human IgG antibody and chicken IgY antibody mixture.

[0013] In certain embodiments, the concentrations of the mouse anti-human IgG antibody and chicken IgY antibody are 0.04-0.06 mg / mL and 0.003-0.005 mg / mL, respectively.

[0014] In some embodiments, the content of fluorescent microspheres on the conjugate pad is 0.1-0.5 μg / cm.

[0015] In certain embodiments, the microsphere diluent comprises 0.1-0.5% glycine, 1-5% BSA, 1-5% Tween 20, 18-22% sucrose and 0.01-0.1M pH 8.0 Tris-HCl buffer, and the solvent is water.

[0016] By adopting the above technical solution, the reagent card uses immunofluorescence chromatography detection and can be conveniently and quickly applied to the rapid detection of serum, plasma, whole blood and fingertip blood.

[0017] In some embodiments, the spacing between the first detection line, the second detection line, the third detection line and the quality control line is 4-10 mm.

[0018] On the other hand, the present application also provides a Helicobacter pylori typing detection kit containing the fluorescent immunochromatographic detection reagent card.

[0019] In certain embodiments, the kit further comprises a sample diluent, comprising 5-6 g / L Na2HPO4·12H2O, 0.5-0.6 g / L NaH2PO4·2H2O, 8-9 g / L NaCl, 0.4-0.6% Tween 80 by volume, and 0.0.5-0.15% proclin300 by volume.

[0020] On the other hand, the present application also provides the use of the fluorescent immunochromatographic detection reagent card or the kit in Helicobacter pylori antibody typing detection.

[0021] In certain embodiments, the application is to perform the test on whole blood or finger stick blood samples.

[0022] Compared with the prior art, the present application has the following technical effects.

[0023] 1. The Helicobacter pylori antibody typing detection reagent card of the present application adopts capture method and fluorescent immunochromatography to qualitatively detect Helicobacter pylori vacuoling cytotoxin (VacA) antibodies, cytotoxin-associated protein (CagA) antibodies and urease (Urease) antibodies in human serum, plasma, whole blood or fingertip blood samples, and obtains the relative fluorescence signal intensity of various antibodies through immunoassay detection. According to the corresponding reference values ​​of Helicobacter pylori vacuoling cytotoxin antibodies, cytotoxin-associated protein antibodies and urease antibodies, the positive and negative of Helicobacter pylori vacuoling cytotoxin antibodies, cytotoxin-associated protein antibodies and urease antibodies are judged respectively, so as to distinguish Helicobacter pylori type I and type II.

[0024] 2. The use of fluorescent labeling method on the reagent card of this application can improve the sensitivity and repeatability of the product; at the same time, the sample diluent is used to dilute the sample, and the sample is diluted more than 10 times, which can still meet the detection of serum / plasma / whole blood / fingertip blood, thereby improving the detection sensitivity while meeting the detection conditions. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 This is a schematic diagram of the structure of the reagent card for this application.

[0026] Figure 2 This is the product image of the reagent card for this application.

[0027] Figure 3 This is the consistency analysis result between the kit in this application and the product of Berlaut.

[0028] Figure 4 This is the sensitivity test result of the reagent card for this application.

[0029] Figure 5 This is the repeatability test result of the reagent card for this application.

[0030] Figure 6 This is the fitted linear curve diagram of the reagent card Urease in this application.

[0031] Figure 7 This is the fitted linear curve diagram of the reagent card CagA of this application.

[0032] Figure 8 This is the fitted linear curve diagram of the reagent card VacA of this application.

[0033] Fig. 9 These are the test results of serum, plasma, whole blood and fingertip blood of the Urease kit of this application.

[0034] Fig.10 These are the test results of serum, plasma, whole blood and fingertip blood of CagA in the kit of this application.

[0035] Fig.11These are the test results of VacA in serum, plasma, whole blood and fingertip blood of the kit of this application.

[0036] Fig.12 This is the anti-interference analysis result of jaundice, hemolysis and lipemia samples of the reagent card in this application.

[0037] Fig.13 These are the anti-interference analysis results of Candida albicans, Enterococcus faecalis, IgG, and IgM samples of the reagent card applied for this application.

[0038] Fig.14 The results of typing test on 16 HP positive serum samples randomly tested using the reagent card in this application. DETAILED DESCRIPTION

[0039] The present application is further described in detail below in conjunction with the accompanying drawings and examples. It is particularly noted that if no specific conditions are specified in the following examples, the experiments are carried out under conventional conditions or conditions recommended by the manufacturer. Unless otherwise specified, the raw materials used in the following examples can all be sourced from common commercial sources.

[0040] Example Preparation of Helicobacter pylori Antibody Typing Immunochromatographic Reagent Card 1. Preparation of coating membrane Helicobacter pylori vacuolar cytotoxin antigen, cytotoxin-associated protein antigen, urease A antigen and urease B antigen were diluted to 0.1 mg / mL with 0.02 M pH 7.4 PBS buffer, and urease A and urease B antigens were mixed at a volume ratio of 1:1 to form an antigen mixture. The goat anti-chicken IgY (purchased from Wuhan Yanda Biotechnology Co., Ltd.) in the quality control area (C) was diluted to 0.5 mg / mL, and the speed was 50 mm / s, the dosage was 1 μL / cm, and they were drawn in parallel on the nitrocellulose membrane as the test line and the quality control line for coating. The quality control line and the test line were separated by 5 mm and placed in an oven and dried at 45°C for 96 h.

[0041] 2. Preparation of conjugate pad (1) Preparation of chicken IgY fluorescent conjugate: Take 1 mL of fluorescent microspheres and activate them with 1-ethyl-(3-dimethylaminopropyl)carbodiimide and N-hydroxysuccinimide for 40 min; wash them 5 times; add 0.4 mg of antibody chicken IgY (purchased from Wuhan Yanda Biotechnology Co., Ltd.), mix well and cross-link for 3 h; add BSA, mix well and block for 1 h; wash them 3 times; and store them in preservation solution.

[0042] (2) Preparation of mouse anti-human IgG fluorescent conjugate: Take 1 mL of fluorescent microspheres and activate them with 1-ethyl-(3-dimethylaminopropyl) carbodiimide and N-hydroxysuccinimide for 40 min; wash 5 times; add 0.4 mg anti-mouse anti-human IgG (purchased from Feipeng Biotechnology Co., Ltd.), mix well and cross-link for 3 h; add BSA, mix well and block for 1 h; wash 3 times; and store in preservation solution.

[0043] (3) On the conjugate pad, the fluorescent microsphere-labeled mouse anti-human IgG antibody and chicken IgY antibody were diluted with a microsphere diluent, wherein the microsphere diluent comprises: 0.3% glycine, 3% casein sodium salt, 2% Tween 20, 20% sucrose and 0.05M pH 8.0 Tris-HCl buffer, and the solvent is water; the diluted mixed solution is evenly sprayed on a line at a dosage of 4 μL / cm, and placed in an oven at 45±2°C for 46 hours. The content of fluorescent microspheres on the conjugate pad is 0.2 μg / cm.

[0044] 3. Preparation of sample pad The prepared 0.02M Tris solution of pH 8.0 is evenly coated on the blank glass fiber, and the laid sample pad is placed in a drying oven or drying room for drying, the drying temperature is 45°C, the humidity is ≤30%, and the drying time is 20 hours. The 0.02M Tris solution contains: 0.8% PVA, 2% casein sodium salt, 2.5% Tween-20, 0.1% Proclin-300, and 0.1mg / mL anti-erythrocyte antibody.

[0045] 4. Reagent card assembly The sample pad, conjugate pad, nitrocellulose membrane and absorbent paper are sequentially attached to the coated plate in overlapping order to obtain a reagent card, such as Figure 1 As shown, the prepared reagent card is fixed on the plastic base card, the surface of the reagent card is pressed tightly with the card surface, and the card surface is reserved with a sample addition hole and an observation window at the portion corresponding to the sample pad and the nitrocellulose membrane, as shown in FIG. Figure 2 shown.

[0046] Performance Evaluation Consistency of effect example 1 with the product of Shenzhen Broute (Shenzhen Broute Helicobacter pylori antibody typing test kit (Western blotting method)) 133 random adult serum samples were collected from the hospital, and the Helicobacter pylori antibody typing detection kit (fluorescent immunochromatography) prepared in the embodiment was used to perform antibody detection on its supporting instrument. At the same time, the reference reagent Berlot Helicobacter pylori detection kit (immunoblotting) was used to perform the same detection, and the consistency of the detection results of the detection reagent prepared in the embodiment of the present application and the Berlot reagent was analyzed respectively. Among them, the Berlot product detects four antibodies, UreA, UreB, CagA and CagA, respectively. In this result, the positive result of any one of UreA and UreB antibodies is judged as the positive result of Urease antibody.

[0047] The results are as follows Figure 3 As shown, "positive detection result" refers to the detection result of the test kit prepared in the embodiment of the present application. In Table 2-1, Table 2-2 and Table 2-3, the results are analyzed by kappa, and the total compliance rate of urease is 96.24%, K=0.92; the total compliance rate of caga is 95.49%, K=0.91; the total compliance rate of vaca is 96.24%, K=0.92, indicating that the test results of Urease, CagA and CagA antibody prepared in the embodiment of the present application (positive Helicobacter pylori antibody typing detection kit) (immunofluorescence method) are highly consistent with the detection results of Shenzhen Berlaut Helicobacter pylori antibody typing detection kit (immunoblotting method).

[0048] Effect Example 2 Sensitivity Detection One Helicobacter pylori type I positive sample was selected and graded diluted using the sample diluent. The Helicobacter pylori antibody typing detection kit (fluorescent immunochromatography) prepared in the example was used to perform antibody detection on its supporting instrument. At the same time, the reference reagent New Sai Ya Helicobacter pylori (HP) detection kit (immunoblotting) was used to perform the same detection. The sensitivity of the detection results of the detection reagent prepared in the example of the present application and the New Sai Ya reagent were analyzed respectively. The results are as follows: Figure 4-5 As shown, the reagent card of this embodiment can still maintain a high response when the sample is diluted 10 times, and can still maintain a positive detection result when the sample is diluted 80 times by the sample diluent (the components are shown in Table 1), which significantly increases the sensitivity of the detection.

[0049] Table 1 Sample dilution components Reagents Sample diluent <![CDATA[Na2HPO4·12H2O]]> 5.80g <![CDATA[NaH2PO4·2H2O]]> 0.59g NaCl 8.50g Twain 80 5.00ml Proclin300 1.00ml Pure water Make up to 1L Effect Example 3: Linear Detection 10 appropriate samples were selected, and the colorimetric distinction and assignment were performed using the Berlaut product (as shown in Table 2). The kit of this application was used for detection, and the linear curves of Urease, CagA and VacA were fitted respectively. The results are as follows Figure 6-8 As shown, the linear correlation coefficients R of Urease, CagA and VacA were 0.9993, 0.9993 and 0.9989, respectively.

[0050] Table 2 Berlot assigned chromaticity Effect Example 4 Repeatability Test Three samples with low, medium and high positive rates of Helicobacter pylori type I were selected and tested 10 times respectively. The precision of the response values ​​of Urease, CagA and VacA were calculated respectively. The results are as follows: Fig. 9 As shown, the accuracy of the detection results of three antibodies, Urease, CagA and VacA, in the three samples with medium and high positive was within 10%.

[0051] Effect Example 5 Correlation between serum, plasma, whole blood and fingertip blood 83 random adult serum, whole blood and fingertip blood samples were collected, and antibody detection was performed on the matching instrument using the kit prepared in the embodiment of the present application (a positive Helicobacter pylori antibody typing detection kit), and the test results of serum, plasma, whole blood and fingertip blood were analyzed.

[0052] The results are as follows Figure 9-11 As shown; Urease( Fig. 9 )、CagA( Fig.10 )、VacA( Fig.11 )'s serum, plasma, whole blood and fingertip blood test results were 1:1, and the correlation coefficients R were 0.9874, 0.9884 and 0.9843, respectively; no bleeding was found during the whole blood and fingertip blood testing.

[0053] Effect Example 6 Interference / Crossover Test A total of 21 samples were tested for 3 cases of icterus, hemolysis, lipemia, and the addition of Candida albicans, Enterococcus faecalis, IgG, and IgM, and then anti-interference analysis was performed. The results are as follows Fig.12 , 13 As shown, for samples of jaundice, hemolysis, and lipemia, the test results of the product of this application are consistent with those of the control reagent Blaute product; the test results of Candida albicans, Enterococcus faecalis, IgG, and IgM samples before and after addition are consistent using this product. The kit prepared in the embodiment of this application has strong anti-interference ability.

[0054] Effect Example 7 Randomly test 16 HP positive serum samples for typing test, the results are as follows Fig.14 As shown, among the 16 positive samples, 5 samples were positive for Urease antibodies, but negative for CagA and VacA antibodies, indicating type 2 Helicobacter pylori infection; the remaining 11 samples were positive for Urease, CagA and VacA antibodies, indicating type 1 Helicobacter pylori infection.

[0055] This specific embodiment is merely an explanation of the present application and is not a limitation of the present application. After reading this specification, those skilled in the art may make creative modifications to the present embodiment as needed, but as long as they are within the scope of the claims of the present application, they are protected by the patent law.

Claims

1. A Helicobacter pylori typing fluorescent immunochromatographic detection reagent card, characterized in that: It includes a coating plate, a sample pad, a conjugation pad, a nitrocellulose membrane and absorbent paper, wherein the sample pad, the conjugation pad, the nitrocellulose membrane and the absorbent paper are overlapped in sequence and pasted on the coating plate; A first detection line sprayed with VacA antigen, a second detection line sprayed with CagA antigen, a third detection line sprayed with a mixed solution of UreA and Ure B antigens, and a quality control line sprayed with goat anti-chicken IgY antibody are arranged in parallel from one end of the conjugation pad of the nitrocellulose membrane to one end of the absorbent paper, wherein the first detection line, the second detection line, the third detection line and the quality control line are parallel to the overlap seam; and a mixed solution of mouse anti-human IgG antibody and chicken IgY antibody marked with fluorescent microspheres is sprayed on the conjugation pad.

2. The fluorescent immunochromatographic detection reagent card according to claim 1, characterized in that: The amino acid sequence of the VacA antigen is shown in SEQ ID NO.1, the amino acid sequence of the CagA antigen is shown in SEQ ID NO.2, the amino acid sequence of the Ure A antigen is shown in SEQ ID NO.3, and the amino acid sequence of the Ure B antigen is shown in SEQ ID NO.

4.

3. The fluorescent immunochromatographic detection reagent card according to claim 2, characterized in that: The concentration of the VacA antigen, CagA antigen, UreA antigen and UreB antigen is 0.05-0.5 mg / mL, preferably 0.1-0.3 mg / mL; the concentration of the goat anti-chicken IgY antibody is 0.1-1 mg / mL, preferably 0.3-0.8 mg / mL.

4. The fluorescent immunochromatographic detection reagent card according to claim 1, characterized in that: The preparation method of the fluorescent microsphere-labeled mouse anti-human IgG antibody and chicken IgY antibody mixed solution is as follows: mouse anti-human IgG antibody and chicken IgY antibody are respectively labeled with fluorescent microspheres, then diluted with microsphere diluent, and mixed at a volume ratio of 1:20 to obtain the fluorescent microsphere-labeled mouse anti-human IgG antibody and chicken IgY antibody mixed solution.

5. The fluorescent immunochromatographic detection reagent card according to claim 4, characterized in that: The concentrations of the mouse anti-human IgG antibody and the chicken IgY antibody are 0.04-0.06 mg / mL and 0.003-0.005 mg / mL respectively.

6. The fluorescent immunochromatographic detection reagent card according to claim 4, characterized in that: The content of the fluorescent microspheres on the binding pad is 0.1-0.5 μg / cm.

7. The fluorescent immunochromatographic detection reagent card according to claim 4, characterized in that: The microsphere diluent comprises 0.1-0.5% glycine, 1-5% BSA, 1-5% Tween 20, 18-22% sucrose and 0.01-0.1 M pH 8.0 Tris-HCl buffer, and the solvent is water.

8. A Helicobacter pylori typing detection kit containing the fluorescent immunochromatographic detection reagent card according to any one of claims 1 to 7, characterized in that: The kit also includes a sample diluent, which includes 5-6 g / L Na2HPO4·12H2O, 0.5-0.6 g / L NaH2PO4·2H2O, 8-9 g / L NaCl, 0.4-0.6% Tween 80 by volume, and 0.0.5-0.15% proclin300 by volume.

9. Use of the fluorescent immunochromatographic detection reagent card according to any one of claims 1 to 7 or the kit according to claim 8 in the detection of Helicobacter pylori antibody typing.

10. The use according to claim 9, characterized in that: The application is to test samples such as serum / plasma / whole blood / fingertip blood.

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