Establishment, Identification And Preliminary Application Of Hybridoma Cell Linces Secreting Monoclonal Antibodies Against Swine Influenza Virues (HN Strian)

Hybridomas secreting monoclonal antibodies (McAbs)specific for Swine influenza virus were produced by fusion of SP2/0 myelomas cells with spleen cells from BALB/C mice immunized with purified SIV.Wells containing hybridomas secreting immunoglobulin G (IgG1) antibodies against SIVwere specifically identified by an indiect enzyme-linked immunosorbent assay(ELISA),and one ELISA-positive hybridomas were selected and cloned forth by limiting dilution. and named 5B4,which could secrete McAb stably.The isotype of the McAb was IgG1,and the average number of chromosome of the hybridoma cell lines was 96.8.The ELISA titer of cell supernate and ascites were respectively 1:1024and 1:51200.The McAb did n't cross-react with PRV,PRRSV,HCV,PPV et al,this demonstrated it has good specifity.The 5B4cell lines can steadly secreted McAb after subcultured twenty-one times .Indirect –ELISA and HI test for detecting SIV were developed by using 5B4 strain monoclonal antibody. The best optimal coating concentration of McAb was 2.95μg/ml,and working concentration of rabbit-anti-SIV positive serum was 1:400.The sensitivity of the double decker Sandwhich-ELISA was 2.43μg/ml. No cross-reaction were dected with major pathogenic microbe,and the high positive coincidence rate with virus isolation assay .The results demonstrated that positive rates sequence is bronchia,trachea,lung,tonsil, scrofula of pigs infected by SIV. And the dipping things obtained by nasal cavity swab that can be dected under no any special disposed, by the double decker sandwhich-ELISA .