The Generation And Identcfication And Application Of Monoclonal Antibodies Against CSFV-SM Strain

In this study ten monoclonal antibodies designated as 2F4,2F7,2E10,2B2, 4G5H2, 5F11, 5C9B1,, 4A1C9, 3C4, 5H11 against classical swine fever virus Shimen strain(CSFV-SM) were generated by immunization of BALB/C mice with purified CSFV-SM strain, using monoclonal antibody technique. The antibody of the supernatant and ascitic fluid from seven hybridomas were detected by indirect ELISA.The titers respectively ranges 1:160 to 1:640 and 1:800 to 1:6400. They can still stably excreted McAb after being stored at -80C.for three months. 2F7, 2E10, 2B2, 4G5H2, 5F11, 5C9B1, and 4A1C9 can reacte with CSFV-SM strain prepared in PK15 cell in 96 microplates using indirect immunofluorescent assay(IFA), especially 4G5H2 has strong fluorescent reaction , it has a titer of 1:1000. The reaction of McAb With CSFV E2 protein by indirect ELISA indicated that 2F7, 2E10, 2B2, 4G5H2 are McAb against CSFV E2 protion. 2F7, 2E10, 2B2, 4G5H2, 5F11, 5C9B, and 4A1C9 could still react with CSFV-SM strsin treated by SDS, indicating the antigen epitope of McAbs are linear epitope.The 4G5H2 could react with CSFV-SM strain, wuzhou strain , BoBai strain, BeiHai strain and vaccine strain prepared in PK15 cell in 96 microplate by immunoenzymatic essay, demonstrating 4G5H2 can recognize all these CSFV strain. 4G5H2 McAb was boated in the 96 well microplates. 22 samples from different areas were detected by the antigen capture assay. 15 clinic samples were positive of CSFV. The result is in agreement with that of PCR based technique.