Preparation of Monoclonal Antibodies Against Peste des petits ruminant virus(PPRV) Nucleoprotein and Establishment of a Competitive ELISA for the Detection of PPRV Antibodies
Abstract：Peste des petits ruminants(PPR) is an acute and highly contagious disease caused by Peste des petits ruminants virus (PPRV). For the detection of IgG against PPRV in sera, BALB/c mice(Mus musculus) were immunized with purified recombinant nucleoprotein of PPRV expressed in eukaryotic expression system. The hybridomas were screened by indirect ELISA using purified recombinant nucleoprotein of PPRV expressed in prokaryotic expression system as coating antigen. Eight monoclonal antibodies(mAbs) against PPRV N protein were filtered and the best was 1D5. Isotype of the 1D5 was IgG2b, and the titer of theascites induced by 1D5 was 1∶106. A competitive ELlSA(c-ELISA) for the detection of antibodies agaist PPRV was established using rpET-PPRN protein as coating antigen and mAb 1D5 as detecting antibody. The c-ELISA could differentiate rinderpest (RP) and PPR positive sera, and was sensitive and specific. According to the test results of 129 goat(Capre hircus) serum samples, the c-ELISA established in this study had a high coincidence rate of 96.9% compared with the c-ELISA kit supplied by CIRAD-EMVT. In this study a c-ELISA to detect IgG against PPRV in sera specifically is established, it’s significant for diagnosis and control of the disease and monitoring the effectiveness of vaccine.
