Influenza A virus and influenza B virus sandwich method detection reagent and application thereof

Abstract

The invention provides an influenza A / B virus sandwich method detection reagent. The influenza A / B virus sandwich method detection reagent 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 of the 19C7 antibody 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 of the 27H4 antibody is SEQ ID NO: 10; the pair of influenza B virus antibodies comprises a labeled antibody 10G11 and a coated antibody 23E2, the heavy chain amino acid sequence of the 10G11 antibody is SEQ ID NO: 17, and the light chain amino acid sequence of the 10G11 antibody 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 of the 23E2 antibody is SEQ ID NO: 26. The sandwich method detection reagent has extremely high specificity and sensitivity.

Description

TECHNICAL FIELD

[0001] The present application relates to a sandwich method detection reagent for influenza A and B virus and its application, belonging to the field of antibody technology. BACKGROUND

[0002] Influenza virus belongs to RNA virus, and belongs to orthomyxoviridae. According to the antigenic difference of nucleoprotein (nucleoprotein, NP) and matrix protein (matrix protein, MP) of the virus, influenza virus can be divided into four types: A, B, C and D. Among them, the type mainly pathogenic to human is influenza A virus, which is the most virulent and easy to mutate; the second is influenza B virus. Therefore, the detection research of influenza A and B virus has great clinical significance.

[0003] Influenza A virus has a wide host range and complex serotypes, and can obtain cross-host transmission ability through genetic recombination and adaptive mutation. Human has no long-lasting immunity to influenza A virus, so people of all ages are generally susceptible, and it is the main pathogen causing pandemic influenza. According to the combination of outer membrane hemagglutinin (HA) and neuraminidase (NA), it can be further divided into several subtypes, and there are 16 H subtypes (H1 - H16) and 9 N subtypes (N1 - N9), and there have been reports of human infection with H1, H3, H5, H7 and other subtypes. Influenza A virus infection in patients is often accompanied by fever, and the body temperature can reach 39℃, and even more than 39℃.

[0004] Influenza B virus is mainly spread in a small range, and there is no subtype division at present, and the main cause of human infection is influenza B Yamagata or influenza B Victoria. In 15 countries in the Asia-Pacific region, the confirmed rate of influenza B virus in confirmed cases of influenza was 0 - 92% per month. Children, the elderly and other specific populations are susceptible to influenza B virus, and are easy to cause complications, and the burden caused to society even exceeds that of influenza A virus.

[0005] Currently, there are three main methods for detecting influenza viruses. The first method is isolation and culture, which is the gold standard for detecting influenza viruses, but the operation is complex, the detection time is long, and the detection cycle is about 14 days, which is difficult to widely carry out in clinical practice. The second method is nucleic acid detection, which has high sensitivity, and compared with the isolation and culture method, the experimental time is greatly shortened, and the clinical application is also more widely. However, this method has high requirements for professional operation, needs precise and expensive instruments, has high cost, and the experimental time also needs 4-6 hours. The third method is rapid antigen detection, which is convenient and fast, does not need any instrument, has simple operation, can be used for self-test, and only needs 10-15 minutes, which is especially suitable for suspected patients self-test screening, special department screening (such as blood transfusion, preoperative, emergency department, etc. in hospital), disease control system screening and entry-exit inspection, etc. However, the specificity and sensitivity of most antigen detection reagents on the market are insufficient, which is difficult to meet the actual application requirements. SUMMARY The application provides an influenza A and B virus sandwich detection reagent and application thereof, which can effectively solve the above problems.

[0006] An influenza A and B virus sandwich detection reagent 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, wherein: The amino acid sequences of the heavy chain CDR1 sequence, the CDR2 sequence and the CDR3 sequence of the 19C7 antibody are shown in SEQ ID NO: 3-5, and the amino acid sequences of the light chain CDR1 sequence, the CDR2 sequence and the CDR3 sequence are shown in SEQ ID NO: 6-8; the amino acid sequences of the heavy chain CDR1 sequence, the CDR2 sequence and the CDR3 sequence of the 27H4 antibody are shown in SEQ ID NO: 11-13, and the amino acid sequences of the light chain CDR1 sequence, the CDR2 sequence and the CDR3 sequence 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 sequence, the CDR2 sequence and the CDR3 sequence of the 10G11 antibody are shown in SEQ ID NO: 19-21, and the amino acid sequences of the light chain CDR1 sequence, the CDR2 sequence and the CDR3 sequence are shown in SEQ ID NO: 22-24; the amino acid sequences of the heavy chain CDR1 sequence, the CDR2 sequence and the CDR3 sequence of the 23E2 antibody are shown in SEQ ID NO: 27-29, and the amino acid sequences of the light chain CDR1 sequence, the CDR2 sequence and the CDR3 sequence are shown in SEQ ID NO: 30-32. 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.

[0007] In some embodiments, the labeled antibody is coupled with a labeled microsphere, and the coating antibody is fixed on a nitrocellulose membrane to form a colloidal gold sandwich assay system.

[0008] In some embodiments, the coating concentration of the coating antibody 27H4 is 0.35-0.45 mg / mL, and the coating concentration of the coating antibody 23E2 is 0.45-0.55 mg / mL.

[0009] In some embodiments, the spraying amount of the labeled antibody 19C7 is 0.5-1.0 μg per centimeter of the conjugate pad, and the spraying amount of the labeled antibody 10G11 is 0.5-1.5 μg per centimeter of the conjugate pad.

[0010] In some embodiments, the quality control line C of the test strip is coated with a goat anti-mouse IgG polyclonal antibody.

[0011] In some embodiments, the drying condition of the nitrocellulose membrane is 50-60℃ for 3-5 days.

[0012] An influenza A and B virus sandwich assay kit comprises the influenza A and B virus sandwich assay test strip.

[0013] In some embodiments, the influenza A and B virus sandwich assay kit further comprises a sample processing buffer.

[0014] The beneficial effects of the present application are: The present application relates to a sandwich method detection reagent for influenza A and B virus, which is specially used for efficient detection of influenza A and B virus antigens, and has the characteristics of high specificity and sensitivity. Specifically, the detection reagent has a minimum detection limit of 200 TCID50 / mL in the detection of influenza A and B virus, ensuring accurate detection even at low virus concentration. In addition, the detection reagent shows excellent detection ability for various influenza A virus strains (including but not limited to H1N1, H3N2, H5N1, H7N9) and influenza B virus strains (such as Victoria and Yamagata), and does not cross-react between virus strains, ensuring the accuracy and reliability of the detection results.

[0015] The detection reagent of the present application does not cross-react with respiratory syncytial virus, mycoplasma pneumoniae, chlamydia pneumoniae, novel coronavirus and other common respiratory pathogens during detection, thereby effectively avoiding the occurrence of false positive results. In addition, the detection results of the detection reagent are not interfered by blood components, drug components or various chemical reagents, ensuring the stability of the detection process and the accuracy of the results, and providing strong technical support for clinical diagnosis and epidemic prevention. DETAILED DESCRIPTION

[0016] To make the purpose, technical scheme and advantages of the embodiments of the present application clearer, the technical scheme of the embodiments of the present application will be described clearly and completely below in combination with the embodiments of the present application. Obviously, the described embodiments are part of the embodiments of the present application, rather than all the embodiments of the present application. Based on the embodiments of the present application, all other embodiments obtained by those skilled in the art without creative labor fall within the scope of protection of the present application. Therefore, the following detailed description of the embodiments of the present application is not intended to limit the scope of the claimed application, but only represents selected embodiments of the present application.

[0017] Example 1 Antibody Preparation The embodiments of the present application prepared two monoclonal antibodies of influenza A virus antibodies, 19C7 and 27H4, and two monoclonal antibodies of influenza B virus antibodies, 10G11 and 23E2.

[0018] 19C7 and 27H4 recognize influenza virus N protein. 10G11 and 23E2 recognize influenza B virus N protein.

[0019] The preparation method is as follows: the mice are immunized multiple times with influenza A and B virus antigens, and then positive mice are screened by ELISA, and their spleen cells are fused to prepare hybridoma cells, and further screening is performed to obtain monoclonal hybridoma cells capable of secreting high-affinity active and high-neutralizing activity antibodies, and then influenza A and B virus monoclonal antibodies are prepared, respectively.

[0020] The mouse immunization method, ELISA screening method, hybridoma cell fusion, screening and culture method are well known to those skilled in the art. The specific information is as follows: The amino acid sequence of the monoclonal antibody 19C7 is as follows: The antibody heavy chain sequence is: EVQLQQSGPELVKPGASVKMSCTASGYTFTSYTVHWVRQKPGQGLEWIGYILPYNDGTKYNEKFKGKATLTSDRSSSTAYMELNSLTSEDSAVFYCARWGWDGFDYWGQGTTLTISSASTKGPSVYPLAPGSAAQTNSMVTLGCLVKGYFPEPVTVTWNSGSLSSGVHTFPAVLQSDLYTLSSSVTVPSSTWPSQTVTCNVAHPASSTKVDKKIVPRDCGCKPCICTVPEVSSVFIFPPKPKDVLTITLTPKVTCVVVDISKDDPEVQFSWFVDDVEVHTAQTKPREEQINSTFRSVSELPIMHQDWLNGKEFKCRVNSAAFPAPIEKTISKTKGRPKAPQVYTIPPPKEQMAKDKVSLTCMITNFFPEDITVEWQWNGQPAENYKNTQPIMDTDGSYFVYSKLNVQKSNWEAGNTFTCSVLHEGLHNHHTEKSLSHSPGK (SEQ ID NO: 1) The antibody light chain sequence is: DVVMTQTPLSLPVSLGDQASISCRSSQSFVHSNGNTYLHWYLQKPGQSPKLLIYKVSNRFSGVPDRFSGSGSGTDFTLRISRVEAEDLGVYFCSQSAHIPPTFGGGTKLEIKRTVAAPSVSIFPPSSEQLTSGGASVVCFLNNFYPKDINVKWKIDGSERQNGVLNSWTDQDSKDSTYSMSSTLTLTKDEYERHNSYTCEATHKTSTSPIVKSFNRNEC (SEQ ID NO: 2) H-CDR1 : SYTVH (SEQ ID NO: 3) H-CDR2: YILPYNDGTKYNEKFKG (SEQ ID NO: 4) H-CDR3: WGWDGFDY (SEQ ID NO: 5) L-CDR1 : RSSQSFVHSNGNTYLH (SEQ ID NO: 6) L-CDR2: KVSNRFS (SEQ ID NO: 7) L-CDR3: SQSAHIPPT (SEQ ID NO: 8) The amino acid sequence of monoclonal antibody 27H4 is as follows: The antibody heavy chain sequence is: QVQLQQPGAELVKPGASVKLSCKTSGYIFTSYWMHWLKQRPGHGLEWIGEINPSNGRTNYNEKFKTKATLTVDKSSSTAYIQLSSLRSEDSAVYYCARDDYDGDWGQGTTLTVSSASTKGPSVYPLAPGSAAQTNSMVTLGCLVKGYFPEPVTVTWNSGSLSSGVHTFPAVLQSDLYTLSSSVTVPSSTWPSQTVTCNVAHPASSTKVDKKIVPRDCGCKPCICTVPEVSSVFIFPPKPKDVLTITLTPKVTCVVVDISKDDPEVQFSWFVDDVEVHTAQTKPREEQINSTFRSVSELPIMHQDWLNGKEFKCRVNSAAFPAPIEKTISKTKGRPKAPQVYTIPPPKEQMAKDKVSLTCMITNFFPEDITVEWQWNGQPAENYKNTQPIMDTDGSYFVYSKLNVQKSNWEAGNTFTCSVLHEGLHNHHTEKSLSHSPGK (SEQ ID NO: 9) The antibody light chain sequence is: DILMTQSPSSMSVSLGDTVSITCHASQGISSNIGWLQQKPGKSFKGLIYHGTNLEDGVPSRFSGSGSGADYSLTISSLEFEDFADYYCVHYAQFPYTFGGGTKLEIKRTVAAP SVSIFPPSSEQLTSGGASVVCFLNNFYPKDINVKWKIDGSERQNGVLNSWTDQDSKDSTYSMSSTLTLTKDEYERHNSYTCEATHKTSTSPIVKSFNRNEC(SEQIDNO:10) H-CDR1: SYWMH (SEQ ID NO:11) H-CDR2: EINPSNGRTNYNEKFKT (SEQ ID NO:12) H-CDR3:DDYDGD (SEQ ID NO:13) L-CDR1: HASQGISSNIG (SEQ ID NO:14) L-CDR2: HGTNLED (SEQ ID NO:15) L-CDR3: VHYAQFPYT (SEQ ID NO:16) The amino acid sequence of monoclonal antibody 10G11 is as follows: The antibody heavy chain sequence is as follows: QVNIQNSGTEIAKPGGSVKFTCRASGYTWTKFTVWWIYERPAQGIEWIFTDFGSVWKEYQERLWEKATLSADRSGATAYYQLNSVMSEDGISYCARHSWAFLSTETWGNGTTVTVASAKTTPPSVYPLAPGSAAQTNSMVTLGCLVKGYFPEPVTVTWNSGSLSSGVHTFPAVLQSDLYTLSSSVTVPSSTWPSETVTCNVAHPASSTKVDKKIVPRDCGCKPCICTVPEVSSVFIFPPKPKDVLTITLTPKVTCVVVDISKDDPEVQFSWFVDDVEVHTAQTQPREEQFNSTFRSVSELPIMHQDWLNGKEFKCRVNSAAFPAPIEKTISKTKGRPKAPQVYTIPPPKEQMAKDKVSLTCMITDFFPEDITVEWQWNGQPAENYKNTQPIMDTDGSYFVYSKLNVQKSNWEAGNTFTCSVLHEGLHNHHTEKSLSHSPG (SEQ ID NO: 17) The antibody light chain sequence is: DIFLSNSPAIHSGSPGDKVSMTCGGASAFTWLKWYNQHSGTSVKRWIYESGRVGTGIPVKFSASSSGTSYTLTISSHDAEDAGTYYCENYTGQVWSFGGATKIEIKRADAAPTVSIFPPSSEQLTSGGASVVCFLNNFYPKDINVKWKIDGSERQNGVLNSWTDQDSKDSTYSMSSTLTLTKDEYERHNSYTCEATHKTSTSPIVKSFNRNEC (SEQ ID NO: 18) Heavy chain variable region CDR1: KFTVW (SEQ ID NO: 19) Heavy chain variable region CDR2: TDFGSVWKEYQERLWE (SEQ ID NO: 20) Heavy chain variable region CDR3: HSWAFLSTET (SEQ ID NO: 21) Light chain variable region CDR1: GGASAFTWLK (SEQ ID NO: 22) Light chain variable region CDR2: ESGRVGT (SEQ ID NO: 23) Light chain variable region CDR3: ENYTGQVWS (SEQ ID NO: 24) The amino acid sequence of monoclonal antibody 23E2 is as follows: The antibody heavy chain sequence is: QVNIQNSGTEIAKPGSSVKFTCRASGYTWTKFTQWWIYERPVQGIEWIFTDFGTVWKEWQERFWEKATLSAERSGATAYYQLNSVMSEDGLSYCARHSFAFLSTESWGNGMTVTVASAKTTPPSVYPLAPGSAAQTNSMVTLGCLVKGYFPEPVTVTWNSGSLSSGVHTFPAVLQSDLYTLSSSVTVPSSTWPSETVTCNVAHPASSTKVDKKIVPRDCGCKPCICTVPEVSSVFIFPPKPKDVLTITLTPKVTCVVVDISKDDPEVQFSWFVDDVEVHTAQTQPREEQFNSTFRSVSELPIMHQDWLNGKEFKCRVNSAAFPAPIEKTISKTKGRPKAPQVYTIPPPKEQMAKDKVSLTCMITDFFPEDITVEWQWNGQPAENYKNTQPIMDTDGSYFVYSKLNVQKSNWEAGNTFTCSVLHEGLHNHHTEKSLSHSPG (SEQ ID NO: 25) The antibody light chain sequence is: DIFLSESPAIHSSSPGDKVSMTCGGATAFTWIKWYNQRSGTSFKRWIYEHGRVGTGIPVKFSASASGTSYTLTISSWDAEDAGTYYCDNYTGQVFSFGGSTKIEIKRADAAPTVSIFPPSSEQLTSGGASVVCFLNNFYPKDINVKWKIDGSERQNGVLNSWTDQDSKDSTYSMSSTLTLTKDEYERHNSYTCEATHKTSTSPIVKSFNRNEC (SEQ ID NO: 26) Heavy chain variable region CDR1: KFTQW (SEQ ID NO: 27) Heavy chain variable region CDR2: TDFGTVWKEWQERFWE (SEQ ID NO: 28) Heavy chain variable region CDR3: HSFAFLSTES (SEQ ID NO: 29) Light chain variable region CDR1: GGATAFTWIK (SEQ ID NO: 30) Light chain variable region CDR2: EHGRVGT (SEQ ID NO: 31) Light chain variable region CDR3: DNYTGQVFS (SEQ ID NO: 32) Example 2: Preparation of colloidal gold reagent for detecting influenza A and B virus antibodies The coating liquid was used to coat the C-line coating antibody, goat anti-mouse IgG polyclonal antibody, influenza A virus coating antibody 27H4, and influenza B virus coating antibody 23E2, respectively, to a concentration of 0.8 mg / mL, 0.4 mg / mL, and 0.5 mg / mL. With the aid of a membrane marker, each coating liquid was evenly marked on the nitrocellulose membrane at a rate of 1 μL / cm, thereby forming the quality control line C, the influenza A virus detection line T1, and the influenza B virus detection line T2. Subsequently, it was dried at 55°C for 4 days.

[0021] At the same time, the influenza A virus marker antibody 19C7 and the influenza B virus marker antibody 10G11 were coupled and labeled on the latex microspheres and sprayed on the conjugate pad. The amount of influenza A and B viruses used 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. Subsequently, it was placed in a 37°C drying oven for 2.5 hours, and finally assembled into a reagent strip. Example 3: Specificity test of influenza A and B virus antigen reagent detection reagent The virus stock solutions of influenza A viruses H1N1 (ATCC VR-1893), H3N2 (ATCC VR-1679), H5N1 (A / Vietnam / 1194 / 2004), and H7N9 (A / Shanghai / 02 / 2013) and influenza B viruses Victoria (B / ATCC VR-1931) and Yamagata (ATCC VR-1804) from ATCC or the CDC were diluted to 1 × 10 3 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 casein, 0.05 % ProClin-300) were used for specificity testing, and the detection results are shown in Table 1.

[0022]

[0023] As shown in Table 1, the reagent of the present embodiment can detect each type of influenza A and B viruses without cross-reaction, and has high specificity.

[0024] Example 4: Sensitivity test of influenza A and B virus antigen reagent detection reagent The virus stock of influenza A virus H1N1 (ATCC VR-1893), H3N2 (ATCC VR-1679), H5N1 (A / Vietnam / 1194 / 2004), H7N9 (A / Shanghai / 02 / 2013), influenza B virus Victoria lineage (B / ATCC VR-1931), Yamagata lineage (ATCC VR-1804) from ATCC or CDC were gradient diluted for limit of detection determination, each dilution was repeated 5 times, and the positive and negative results were determined. The detection results are shown in Table 2.

[0025]

[0026] The concentrations capable of stably detecting 5 times were selected from Table 2, and two concentrations were diluted by half. The detection was repeated 20 times for each, and the virus level with a 95% positive detection rate was determined as the lowest detection limit. The detection results are shown in Tables 3-8.

[0027]

[0028]

[0029]

[0030]

[0031]

[0032]

[0033] According to Tables 3-8, the lowest detection limit of the reagent for influenza A and B viruses in the embodiment of the present application is 200 TCID50 / mL.

[0034] Example 5: Cross-validation of strains and viruses The different microorganisms in the following table were diluted to the corresponding titers using the sample extraction solution, and the specificity of the reagent of the present application was verified. The related strain and virus titers and detection results are shown in Table 9.

[0035] Table 9

[0036] According to Table 9, the results of the reagent of the present application in detecting the above strains and viruses are all negative, indicating that the specificity of the reagent is very strong, and there is basically no cross-reaction with other microorganisms.

[0037] Example 6: Cross-validation of interferents Different chemical reagents, drugs and whole blood in the following table are diluted to corresponding concentrations using sample extraction solution to verify specificity and anti-interference ability of the reagent of the present application, and the detection results are as shown in Table 10.

[0038] Table 10

[0039] As shown in Table 10, the detection results of the reagent of the present application and the above concentrations of chemical reagents, drugs and whole blood are all negative, which indicates that the specificity of the reagent is very strong, and the detection results are not affected by the above concentrations of chemical reagents, drugs and whole blood.

[0040] The above description is only preferred embodiments of the present application and is not intended to limit the present application. The present application can be variously changed and modified by those skilled in the art. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application shall be included in the protection scope of the present application.

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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