Developed Latex Immuno-chromatographic Assay Of Influenza A Virus

Objectives:To screen monoclonal antibodies against influenza A virus nucleoprotein (NP). To set up an effective and simple method, Latex immunochromatographic assay (LICA), for rapid detection of influenza virus infection.Methods:Protein concentration and purity of six anti-influenza A virus monoclonal antibodies were separately determined by using BCA method and SDS-PAGE method, and then the affinity and specificity of six monoclonal antibodies were identified in the way of the indirect enzyme-linked immunosorbent assay and colloidal gold immunochromatography combination., obtain a pair of antibody.Establish Latex immunochromatographic assay for detection of influenza A virus, sensitivity, specificity, stability and repeatability was identified. LICA assay was used to detect influenza virus A antigen in nasopharyngeal secretions of 349 patients and in the meanwhile, by comparing with PCR and virus cultivation, for verifing sensitivity and specificity of LICA method.Results:The selected best antibody match-AF2 and AF4, was used to establish LICA. Its sensitivity is 5 times for GICA. Application of the kits for detection of influenza A virus for analysis of 349 samples collected from patients, with a coincidence at 59.34% and 100% respectively with the GICA. Comparing with virus cultivation, a coincidence is 63.16% and 93.57% respectively.Conclusion:The method of using LICA to detect rapidly Flu A antigen is simple, convenient and rapid. Its specificity and sensitivity, especially the specificity is good, and the results indicated that there is good appliance of LICA for clinical detection of Influenza viruses.