Generation And Application Of Monoclonal Antibodies Against CSFV Shimen Strain

In this study four monoclonal antibodies designated as AC9, CF8, DG5 and EC9 against classical swine fever virus Shimen strain(CSFV Shimen strain) were generated by immunization of Balb/C mice with purified CSFV Shimen, which multiplied by PK15 cells culture. The hybridomas were inoculated in the mice bellies to produce McAbs. The ascites titer had been measured between 1:2000-1:8000. The McAbs except for DG5 could react with CSFV E2 protein in indirect ELISA, indicating that AC9, CF8 and EC9 are anti-CSFV E2 protein McAbs. AC9, DG5 and EC9 could still react with CSFV Shimen strain treated by SDS, while CF8 react negatively, indicating the structure is essential to the epitope antigenicity of CF8. CF8 and DG5 had strong reactivity CSFV Shimen strain prepared in cells PK15 slides in indirect immunofluorescent assay(IFA), while AC9 and EC9 react weakly in IFA.The comparison of antigen-capture indirect ELJSA(ACI-ELISA) experiments showed the corporation of AC9 and CF8 could doubly enhance reaction sensitivity than AC9 or CF8 alone. Throught chessboard test found out the most suitable dilution of AC9-CF8 and the CSFV postive standar serum respectively is 1:400 and 1:200. The average CSFV titer of samples is 1:32 by indirect ELISA, but 1:4 dilution is used in practice. The results of antigen substitution and antigen blocking experiments showed the ACI-ELISA has highly spectific reactivity with CSFV. At last, the ACI-ELISA was used to test 30 samples, it yielded nearly the same results as PCR.