Influenza a and b virus sandwich assay and use thereof

Abstract

The application provides a sandwich method detection reagent for influenza A and B virus, which comprises a pair of influenza A virus antibodies and a pair of influenza B virus antibodies; the pair of influenza A virus antibodies comprises a labeled antibody 19C7 and a coated antibody 27H4, the heavy chain amino acid sequence of the 19C7 antibody is SEQ ID NO:1, and the light chain amino acid sequence is SEQ ID NO:2; the heavy chain amino acid sequence of the 27H4 antibody is SEQ ID NO:9, and the light chain amino acid sequence is SEQ ID NO:10; the pair of influenza B virus antibodies comprises a labeled antibody 10G11 and a coated antibody 23E2, wherein the heavy chain amino acid sequence of the 10G11 antibody is SEQ ID NO:17, and the light chain amino acid sequence is SEQ ID NO:18; the heavy chain amino acid sequence of the 23E2 antibody is SEQ ID NO:25, and the light chain amino acid sequence is SEQ ID NO:26. The sandwich method detection reagent has very high specificity and sensitivity.

Description

Technical Field

[0001] This invention relates to a sandwich method detection reagent for influenza A and B viruses and its application, belonging to the field of antibody technology. Background Technology

[0002] Influenza viruses are RNA viruses, belonging to the Orthomyxoviridae family. Based on the antigenic differences of their nucleoprotein (NP) and matrix protein (MP), influenza viruses can be classified into four types: type A, type B, type C, and type D. Among them, the type that is pathogenic to humans is mainly type A influenza virus, which is the most virulent and prone to mutation; followed by type B influenza virus. Therefore, research on the detection of type A and type B influenza viruses has significant clinical value.

[0003] Influenza A virus has a wide host range and complex serotypes, and can acquire cross-host transmission capabilities through genetic recombination and adaptive mutation. Humans lack lasting immunity to influenza A virus, making all age groups universally susceptible. It is the most common pathogen causing influenza pandemics. Based on the combination of hemagglutinin (HA) and neuraminidase (NA), it can be further subdivided into several subtypes. Currently, there are 16 H subtypes (H1-H16) and 9 N subtypes (N1-N9), with reports of human infections with subtypes such as H1, H3, H5, and H7. Patients infected with influenza A virus often experience fever, with body temperatures reaching 39°C or even exceeding that.

[0004] Influenza B viruses typically circulate in small, localized areas. Currently, there are no subtype classifications, but the Yamagata or Victoria lineages of influenza B are the primary causes of human infection. In 15 countries in the Asia-Pacific region, the monthly confirmed case rate of influenza B virus ranges from 0% to 92%. Children, the elderly, and other specific populations are particularly susceptible to influenza B virus infection and are prone to complications, placing a greater burden on society than influenza A virus.

[0005] Currently, there are three main methods for detecting influenza viruses. The first is the isolation and culture method, which is the gold standard for influenza virus detection. However, it is complex to operate, time-consuming, and has a testing cycle of approximately 14 days, making it difficult to widely implement in clinical practice. The second is the nucleic acid detection method, which has high sensitivity and significantly shortens the experimental time compared to the isolation and culture method, making it more widely used clinically. However, this method requires a high level of expertise in experimental operation, necessitates sophisticated and expensive instruments, resulting in higher costs, and the experimental time also requires 4-6 hours. The third is the rapid antigen detection method, which is convenient and quick, requires no instruments, is simple to operate, and can be used for self-testing. The test only takes 10-15 minutes and is particularly suitable for self-screening of suspected patients, screening in special departments (such as blood transfusions, pre-operative screening, and emergency departments in hospitals), screening in disease control systems, and entry-exit quarantine scenarios. However, most antigen test reagents on the market lack sufficient specificity and sensitivity, making it difficult to meet practical application needs. Summary of the Invention

[0006] This invention provides a sandwich method detection reagent for influenza A and B viruses and its application, which can effectively solve the above-mentioned problems.

[0007] A sandwich method detection reagent for influenza A and B viruses includes a pair of influenza A virus antibodies and a pair of influenza B virus antibodies;

[0008] The pair of influenza A virus antibodies includes labeled antibody 19C7 and coated antibody 27H4, wherein:

[0009] The amino acid sequences of the heavy chain CDR1, CDR2, and CDR3 sequences of the 19C7 antibody are shown in SEQ ID NO:3-5, and the amino acid sequences of the light chain CDR1, CDR2, and CDR3 sequences are shown in SEQ ID NO:6-8; the amino acid sequences of the heavy chain CDR1, CDR2, and CDR3 sequences of the 27H4 antibody are shown in SEQ ID NO:11-13, and the amino acid sequences of the light chain CDR1, CDR2, and CDR3 sequences are shown in SEQ ID NO:14-16.

[0010] The pair of influenza B virus antibodies comprises a labeled antibody 10G11 and a coated antibody 23E2, wherein: the amino acid sequences of the heavy chain CDR1, CDR2, and CDR3 sequences of the 10G11 antibody are shown in SEQ ID NO:19-21, and the amino acid sequences of the light chain CDR1, CDR2, and CDR3 sequences are shown in SEQ ID NO:22-24; the amino acid sequences of the heavy chain CDR1, CDR2, and CDR3 sequences of the 23E2 antibody are shown in SEQ ID NO:27-29, and the amino acid sequences of the light chain CDR1, CDR2, and CDR3 sequences are shown in SEQ ID NO:30-32.

[0011] In some embodiments, the heavy chain amino acid sequence of the 19C7 antibody is SEQ ID NO:1, and the light chain amino acid sequence is SEQ ID NO:2; the heavy chain amino acid sequence of the 27H4 antibody is SEQ ID NO:9, and the light chain amino acid sequence is SEQ ID NO:10; the heavy chain amino acid sequence of the 10G11 antibody is SEQ ID NO:17, and the light chain amino acid sequence is SEQ ID NO:18; the heavy chain amino acid sequence of the 23E2 antibody is SEQ ID NO:25, and the light chain amino acid sequence is SEQ ID NO:26. A sandwich method test strip for influenza A and B viruses includes the aforementioned detection reagents.

[0012] In some embodiments, the labeled antibody is coupled to labeled microspheres, and the coated antibody is immobilized on a nitrocellulose membrane to form a colloidal gold sandwich detection system.

[0013] In some embodiments, the coating concentration of the coated antibody 27H4 is 0.35-0.45 mg / mL, and the coating concentration of the coated antibody 23E2 is 0.45-0.55 mg / mL.

[0014] In some embodiments, the amount of the labeled antibody 19C7 sprayed is 0.5-1.0 μg / cm conjugate pad, and the amount of the labeled antibody 10G11 sprayed is 0.5-1.5 μg / cm conjugate pad.

[0015] In some embodiments, the control line C of the test strip is coated with goat anti-mouse IgG polyclonal antibody.

[0016] In some embodiments, the nitrocellulose membrane is dried at 50-60°C for 3-5 days.

[0017] A sandwich method detection kit for influenza A and B viruses, comprising the aforementioned sandwich method test strip for influenza A and B viruses.

[0018] In some embodiments, the influenza A and B virus sandwich assay kit further includes a sample processing buffer.

[0019] The beneficial effects of this invention are:

[0020] The sandwich method detection reagent for influenza A and B viruses involved in this invention is specifically designed for the efficient detection of influenza A and B virus antigens, and its significant feature is its extremely high specificity and sensitivity. Specifically, the detection reagent achieves a limit of detection of 200 TCID50 / mL for both influenza A and influenza B viruses, ensuring accurate detection even at low virus concentrations. Furthermore, the reagent demonstrates excellent detection capabilities against various influenza A virus strains (including but not limited to H1N1, H3N2, H5N1, and H7N9) and influenza B virus strains (such as Victoria and Yamagata), and there is no cross-reactivity between the virus strains, ensuring the accuracy and reliability of the detection results.

[0021] The detection reagent described in this invention does not cross-react with respiratory syncytial virus, mycoplasma pneumoniae, chlamydia pneumoniae, novel coronavirus, or other common respiratory pathogens during the detection process, thus effectively avoiding false positive results. Furthermore, the detection results are unaffected by blood components, drug components, or various chemical reagents, ensuring the stability of the detection process and the accuracy of the results, providing strong technical support for clinical diagnosis and epidemic prevention and control. Detailed Implementation

[0022] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention. Therefore, the following detailed description of the embodiments of the present invention is not intended to limit the scope of the claimed invention, but merely to represent selected embodiments of the present invention.

[0023] Example 1 Antibody Preparation

[0024] In this embodiment of the invention, monoclonal antibodies against two types of influenza A virus, namely 19C7 and 27H4, and monoclonal antibodies against two types of influenza B virus, namely 10G11 and 23E2, were prepared.

[0025] 19C7 and 27H4 recognize the N protein of influenza A virus. 10G11 and 23E2 recognize the N protein of influenza B virus.

[0026] The preparation method is as follows: Mice were immunized multiple times with influenza A and B virus antigens, and positive mice were selected by ELISA. Their spleen cells were fused to prepare hybridoma cells. Monoclonal hybridoma cells that could secrete high-affinity and medium-neutralizing antibodies were further screened, and then monoclonal antibodies against influenza A and B viruses were prepared respectively.

[0027] Mouse immunization methods, ELISA screening methods, hybridoma cell fusion, screening and culture methods are well known to those skilled in the art.

[0028] The specific information is as follows:

[0029] The amino acid sequence of monoclonal antibody 19C7 is as follows:

[0030] The antibody heavy chain sequence is as follows:

[0031] EVQLQQSGPELVKPGASVKMSCTASGYTFTSYTVHWVRQKPGQGLEWIGYILPYNDGTKYNEKFKGKATLTSDRSSSTAYMELNSLTSEDSAVFYCARWGWDGFDYWGQGTTL TISSASTKGPSVYPLAPGSAAQTNSMVTLGCLVKGYFPEPVTWNSGSLSSGVHTFPAVLQSDLYTLSSSVTVPSSTWPSQTVTCNVAHPASSTKVDKKIVPRDCGCKPCIC TVPEVSSVFIFPPKPKDVLTITLTPKVTCVVVDISKDDPEVQFSWFVDDVEVHTAQTKPREEQINSTFRSSVSELPIMHQDWLNGKEFKCRVNSAAFPAPIEKTISKTKGRPKA PQVYTIPPPKEQMAKDKVSLTCMITNFFPEDITVEWQWNGQPAENYKNTQPIMDTDGSYFVYSKLNVQKSNWEAGNTFTCSVLHEGLHNHHTEKSLSHSPGK(SEQIDNO:1)

[0032] The antibody light chain sequence is as follows:

[0033] DVVMTQTPLSLPVSLGDQASISCRSSQSFVHSNGNTYLHWYLQKPGQSPKLLIYKVSNRFSGVPDRFSGSGSGTDFTLRISRVEAEDLGVYFCSQSAHIPPTFGGGTKLEIKRTV AAPSVSIFPPSSEQLTSGGASVVCFLNNFYPKDINVKWKIDGSERQNGVLNSWTDQDSKDSTYSMSSTLTLTKDEYERHNSYTCEATHKTSTSPIVKSFNRNEC(SEQIDNO:2)

[0034] H-CDR1: SYTVH (SEQ ID NO:3)

[0035] H-CDR2: YILPYNDGTKYNEKFKG (SEQ ID NO:4)

[0036] H-CDR3: WGWDGFDY (SEQ ID NO:5)

[0037] L-CDR1: RSSQSFVHSNGNTYLH (SEQ ID NO:6)

[0038] L-CDR2: KVSNRFS (SEQ ID NO:7)

[0039] L-CDR3: SQSAHIPPT (SEQ ID NO:8)

[0040] The amino acid sequence of monoclonal antibody 27H4 is as follows:

[0041] The antibody heavy chain sequence is as follows:

[0042] QVQLQQPGAELVKPGASVKLSCKTSGYIFTSYWMHWLKQRPGHGLEWIGEINPSNGRTNYNEKFKTKATLTVDKSSSTAYIQLSSLRSEDSAVYYCARDDYDGDWGQGTTLTVSSASTKGPSVYPLAPGSAAQTNSMVTLGCLVKGYFPEPVTVTWNSGSLSSGVHTFPAVLQSDLYTLSSSVTVPSSTWPSQTVTCNVAHPASSTKVDKKIVPRDCGCKPCICTVPEVSSVFIFPPKPKDVLTITLTPKVTCVVVDISKDDPEVQFSWFVDDVEVHTAQTKPREEQINSTFRSVSELPIMHQDWLNGKEFKCRVNSAAFPAPIEKTISKTKGRPKAPQVYTIPPPKEQMAKDKVSLTCMITNFFPEDITVEWQWNGQPAENYKNTQPIMDTDGSYFVYSKLNVQKSNWEAGNTFTCSVLHEGLHNHHTEKSLSHSPGK(SEQIDNO:9)

[0043] The antibody light chain sequence is:

[0044] DILMTQSPSSMSVSLGDTVSITCHASQGISSNIGWLQQKPGKSFKGLIYHGTNLEDGVPSRFSGSGSGADYSLTISSLEFEDFADYYCVHYAQFPYTFGGGTKLEIKRTVAAPSVSIFPPSSEQLTSGGASVVCFLNNFYPKDINVKWKIDGSERQNGVLNSWTDQDSKDSTYSMSSTLTLTKDEYERHNSYTCEATHKTSTSPIVKSFNRNEC(SEQIDNO:10)

[0045] H-CDR1: SYWMH (SEQ ID NO:11)

[0046] H-CDR2: EINPSNGRTNYNEKFKT (SEQ ID NO:12)

[0047] H-CDR3: DDYDGD (SEQ ID NO:13)

[0048] L-CDR1: HASQGISSNIG (SEQ ID NO:14)

[0049] L-CDR2: HGTNLED (SEQ ID NO:15)

[0050] L-CDR3: VHYAQFPYT (SEQ ID NO:16)

[0051] The amino acid sequence of monoclonal antibody 10G11 is as follows:

[0052] The heavy chain sequence of the antibody is:

[0053] QVNIQNSGTEIAKPGGSVKFTCRASGYTWTKFTVWWIYERPAQGIEWIFTDFGSVWKEYQERLWEKATLSADRSGATAYYQLNSVMSEDGISYCARHSWAFLSTETWGNGTTVTVASAKTTPPSVYPLAPGSAAQTNSMVTLGCLVKGYFPEPVTVTWNSGSLSSGVHTFPAVLQSDLYTLSSSVTVPSSTWPSETVTCNVAHPASSTKVDKKIVPRDCGCKPCICTVPEVSSVFIFPPKPKDVLTITLTPKVTCVVVDISKDDPEVQFSWFVDDVEVHTAQTQPREEQFNSTFRSVSELPIMHQDWLNGKEFKCRVNSAAFPAPIEKTISKTKGRPKAPQVYTIPPPKEQMAKDKVSLTCMITDFFPEDITVEWQWNGQPAENYKNTQPIMDTDGSYFVYSKLNVQKSNWEAGNTFTCSVLHEGLHNHHTEKSLSHSPG (SEQ ID NO:17)

[0054] The light chain sequence of the antibody is:

[0055] DIFLSNSPAIHSGSPGDKVSMTCGGASAFTWLKWYNQHSGTSVKRWIYESGRVGTGIPVKFSASSSGTSYTLTISSHDAEDAGTYYCENYTGQVWSFGGATKIEIKRADAAPTVSIFPPSSEQLTSGGASVVCFLNNFYPKDINVKWKIDGSERQNGVLNSWTDQDSKDSTYSMSSTLTLTKDEYERHNSYTCEATHKTSTSPIVKSFNRNEC (SEQ ID NO:18)

[0056] Heavy chain variable region CDR1: KFTVW (SEQ ID NO:19)

[0057] Heavy chain variable region CDR2: TDFGSVWKEYQERLWE (SEQ ID NO:20)

[0058] Heavy chain variable region CDR3: HSWAFLSTET (SEQ ID NO:21)

[0059] Light chain variable region CDR1: GGASAFTWLK (SEQ ID NO:22)

[0060] Light chain variable region CDR2: ESGRVGT (SEQ ID NO:23)

[0061] Light chain variable region CDR3: ENYTGQVWS (SEQ ID NO:24)

[0062] The amino acid sequence of monoclonal antibody 23E2 is as follows:

[0063] The antibody heavy chain sequence is as follows:

[0064] QVNIQNSGTEIAKPGSSVKFTCRASGYTWTKFTQWWIYERPVQGIEWIFTDFGTVWKEWQERFWEKATLSAERSGATAYYQLNSVMSEDGLSYCARHSFAFLSTESWGNGMTV TVASAKTTPPSVYPLAPGSAAQTNSMVTLGCLVKGYFPEPVTWNSGSLSSGVHTFPAVLQSDLYTLSSSVTVPSSTWPSETVTCNVAHPASSTKVDKKIVPRDCGCKPCIC TVPEVSSVFIFPPKPKDVLTITLTPKVTCVVVDISKDDPEVQFSWFVDDVEVHTAQTQPREEQFNSTFRSSVSELPIMHQDWLNGKEFKCRVNSAAFPAPIEKTISKTKGRPKA PQVYTIPPPKEQMAKDKVSLTCMITDFFPEDITVEWQWNGQPAENYKNTQPIMDTDGSYFVYSKLNVQKSNWEAGNTFTCSVLHEGLHNHHTEKSLSHSPG(SEQIDNO:25)

[0065] The antibody light chain sequence is: DIFLSESPAIHSSSPGDKVSMTCGGATAFTWIKWYNQRSGTSFKRWIYEHGRVGTGIPVKFSASASGTSYTLTISSWDAEDAGTYYCDNYTGQVFSFGGSTKIEIKRA DAAPTVSIFPPSSEQLTSGGASVVCFLNNFYPKDINVKWKIDGSERQNGVLNSWTDQDSKDSTYSMSSSTLTLTKDEYERHNSYTCEATHKTSTSPIVKSFNRNEC(SEQIDNO:26)

[0066] Heavy chain variable region CDR1: KFTQW (SEQ ID NO:27)

[0067] Heavy chain variable region CDR2: TDFGTVWKEWQERFWE (SEQ ID NO:28)

[0068] Heavy chain variable region CDR3: HSFAFLSTES (SEQ ID NO:29)

[0069] Light chain variable region CDR1: GGATAFTWIK (SEQ ID NO:30)

[0070] Light chain variable region CDR2: EHGRVGT (SEQ ID NO:31)

[0071] Light chain variable region CDR3: DNYTGQVFS (SEQ ID NO:32)

[0072] Example 2: Preparation of Antigen Detection Colloidal Gold Reagent for Influenza A and B Virus Antibodies

[0073] The C-line coated antibody (goat anti-mouse IgG polyclonal antibody), influenza A virus coated antibody 27H4, and influenza B virus coated antibody 23E2 were diluted to 0.8 mg / mL, 0.4 mg / mL, and 0.5 mg / mL, respectively, using coating buffer. Using a coating apparatus, each coating buffer was evenly spread onto a nitrocellulose membrane at a rate of 1 μL / cm, thus forming the control line C, the influenza A virus detection line T1, and the influenza B virus detection line T2. The membranes were then dried at 55°C for 4 days.

[0074] Simultaneously, influenza A virus marker antibody 19C7 and influenza B virus marker antibody 10G11 were conjugated onto latex microspheres and sprayed onto the conjugate pads. The dosage of influenza A and influenza B viruses was 0.8 μg of influenza A 19C7 antibody per centimeter of conjugate pad and 1.0 μg of influenza B 10G11 antibody per centimeter of conjugate pad, respectively. Afterwards, the microspheres were placed in a 37°C drying oven for 2.5 hours and finally assembled into test strips.

[0075] Example 3: Specificity test of influenza A and B virus antigen reagent detection reagent

[0076] The stock solutions of influenza A viruses H1N1 (ATCCVR-1893), H3N2 (ATCCVR-1679), H5N1 (A / Vietnam / 1194 / 2004), and H7N9 (A / Shanghai / 02 / 2013) and influenza B viruses Victoria lineage (B / ATCC VR-1931) and Yamagata lineage (ATCC VR-1804) from ATCC or CDC sources were diluted to 1×10⁻⁶. 3 The specificity was tested using TCID50 / mL and sample processing buffer (50 mM Tris-HCl (pH 8.0), 0.5% NaCl, 0.3% Triton X-100, 0.1% Tween-20, 0.2% PVP-40, 0.3% sodium caseinate, 0.05% ProClin-300), and the results are shown in Table 1 below.

[0077]

[0078] As shown in Table 1, the reagents of the present invention can detect various types of influenza A and B viruses without cross-detection, demonstrating high specificity.

[0079] Example 4: Sensitivity Test of Influenza A and B Virus Antigen Reagent Detection Reagent

[0080] The limits of detection were determined by serially diluting the stock solutions of influenza A viruses H1N1 (ATCCVR-1893), H3N2 (ATCCVR-1679), H5N1 (A / Vietnam / 1194 / 2004), and H7N9 (A / Shanghai / 02 / 2013) and influenza B viruses Victoria lineage (B / ATCC VR-1931) and Yamagata lineage (ATCC VR-1804) from ATCC or CDC sources. Each serially diluted virus solution was repeated 5 times, and positive and negative results were detected. The results are shown in Table 2 below.

[0081]

[0082] Five concentrations that could be stably detected were selected from Table 2. These two concentrations were then halved, and each concentration was repeated 20 times. The viral level with a 95% positive detection rate was taken as the limit of detection. The detection results are shown in Table 3-8 below.

[0083]

[0084]

[0085]

[0086]

[0087]

[0088]

[0089] As shown in Tables 3-8, the lowest detection limit for influenza A and B viruses using the reagents implemented in this invention was ultimately determined to be 200 TCID50 / mL.

[0090] Example 5: Cross-verification of strains and viruses

[0091] The different microorganisms in the table below were diluted to the corresponding titers using sample extraction buffer to verify the specificity of the reagent of the present invention. The relevant strains, virus titers and detection results are shown in Table 9 below.

[0092] Table 9

[0093]

[0094] As shown in Table 9, the reagent of this invention tested negative for all the above strains and viruses, indicating that the reagent is highly specific and has virtually no cross-reaction with the other microorganisms mentioned above.

[0095] Example 6: Cross-validation of interfering factors

[0096] The different chemical reagents, drugs, and whole blood in the table below were diluted to the corresponding concentrations using sample extraction buffer to verify the specificity and anti-interference ability of the reagents of the present invention. The test results are shown in Table 10 below.

[0097] Table 10

[0098]

[0099] As shown in Table 10, the test results of the reagent of the present invention were negative for chemical reagents, drugs and whole blood of the above concentrations, indicating that the reagent has high specificity and the test results are not affected by chemical reagents, drugs and whole blood of the above concentrations.

[0100] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention. Various modifications and variations can be made to the invention by those skilled in the art. Any modifications, equivalent substitutions, or improvements made within the spirit and principles of the invention should be included within the scope of protection of the invention.

Claims

1. A sandwich method detection reagent for influenza A and B viruses, characterized in that, This includes a pair of antibodies against influenza A virus and a pair of antibodies against influenza B virus; The pair of influenza A virus antibodies includes labeled antibody 19C7 and coated antibody 27H4, wherein: The amino acid sequences of the heavy chain CDR1, CDR2, and CDR3 sequences of the 19C7 antibody are shown in SEQ ID NO:3-5, and the amino acid sequences of the light chain CDR1, CDR2, and CDR3 sequences are shown in SEQ ID NO:6-8; the amino acid sequences of the heavy chain CDR1, CDR2, and CDR3 sequences of the 27H4 antibody are shown in SEQ ID NO:11-13, and the amino acid sequences of the light chain CDR1, CDR2, and CDR3 sequences are shown in SEQ ID NO:14-16. The pair of influenza B virus antibodies comprises a labeled antibody 10G11 and a coated antibody 23E2, wherein: the amino acid sequences of the heavy chain CDR1, CDR2, and CDR3 sequences of the 10G11 antibody are shown in SEQ ID NO:19-21, and the amino acid sequences of the light chain CDR1, CDR2, and CDR3 sequences are shown in SEQ ID NO:22-24; the amino acid sequences of the heavy chain CDR1, CDR2, and CDR3 sequences of the 23E2 antibody are shown in SEQ ID NO:27-29, and the amino acid sequences of the light chain CDR1, CDR2, and CDR3 sequences are shown in SEQ ID NO:30-32.

2. The sandwich method detection reagent for influenza A and B viruses according to claim 1, characterized in that, The heavy chain amino acid sequence of the 19C7 antibody is SEQ ID NO:1, and the light chain amino acid sequence is SEQ ID NO:2; the heavy chain amino acid sequence of the 27H4 antibody is SEQ ID NO:9, and the light chain amino acid sequence is SEQ ID NO:10; the heavy chain amino acid sequence of the 10G11 antibody is SEQ ID NO:17, and the light chain amino acid sequence is SEQ ID NO:18; the heavy chain amino acid sequence of the 23E2 antibody is SEQ ID NO:25, and the light chain amino acid sequence is SEQ ID NO:

26.

3. A sandwich method test strip for detecting influenza A and B viruses, characterized in that, Includes the detection reagent as described in claim 1 or 2.

4. The influenza A and B virus sandwich test strip according to claim 3, characterized in that, The labeled antibody is coupled to the labeled microspheres, and the coated antibody is immobilized on a nitrocellulose membrane to form a colloidal gold sandwich detection system.

5. The influenza A and B virus sandwich test strip according to claim 4, characterized in that, The coating concentration of the coated antibody 27H4 is 0.35-0.45 mg / mL, and the coating concentration of the coated antibody 23E2 is 0.45-0.55 mg / mL.

6. The influenza A and B virus sandwich test strip according to claim 4, characterized in that, The amount of the labeled antibody 19C7 sprayed onto the conjugate pad is 0.5-1.0 μg / cm, and the amount of the labeled antibody 10G11 sprayed onto the conjugate pad is 0.5-1.5 μg / cm.

7. The influenza A and B virus sandwich test strip according to claim 4, characterized in that, The control line C of the test strip is coated with goat anti-mouse IgG polyclonal antibody.

8. The influenza A and B virus sandwich test strip according to claim 4, characterized in that, The nitrocellulose membrane is dried at 50-60℃ for 3-5 days.

9. A sandwich method detection kit for influenza A and B viruses, characterized in that, Including the sandwich method test strip for influenza A and B viruses as described in any one of claims 3 to 8.

10. The influenza A and B virus sandwich method detection kit according to claim 9, characterized in that, It also includes sample processing buffer.

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