Preparation of monoclonal antibody against peste des petits ruminants virus based on N protein and establishment of a blocking ELISA for detection of antibodies
To establish a rapid antibody detection method for peste des petits ruminants virus(PPRV),in this study,pET-32a-N recombinant plasmid was transformed into E.coli BL21(DE3)competent cells.The recombinant protein was obtained by IPTG-induced expression and purifica-tion,which was used to immunized BALB/c mouse.The monoclonal antibodies against PPRV-N proteins were prepared and HRP-labeled,a PPRV blocking ELISA antibody detection method was established by optimizing the reaction conditions.The method was evaluated to have no cross-reac-tivity with the positive serum of PIV,GTPV,and ORFV;the lowest positive serum could be detec-ted at a dilution of 1∶160;the intra-and inter-batch coefficients of variation(Cv)were less than 10％;and the compliance rate between the method and competitive ELISA kit was 96％by testing 180 clinical serum samples.The results showed that the blocking ELISA has good specificity,sensi-tivity,and reproducibility,and can be used for the detection of PPRV antibodies,which can provide technical support for the evaluation of the immunization effect of PPRV vaccine and the prevention and control of epidemics.
peste des petits ruminantsN proteinmonoclonal antibodyblocking ELISA
NETL
NSTL
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