Preparation and preliminary application of fluorescent antibody against canine distemper virus

To obtain fluoescent antibody against canine distemper virus,monoclonal antibody against N protein of CDV(Onderstepoort strain) was developed by means of the conventional protocol.Ascites was produced using positive colonies and purified by protein G affinity chromatography.Monoclonal antibody was labelled with fluorescein isothiocyanate(FITC) using dialysis methods.Dissociative FITC was removed by Sephadex G-25 column and the F/P value and optimal working dilution of fluorescent antibody(FA) was determined.Absorption test,blocking test and antigen interaction were carried out to identify the specificity of the FA.The results showed six hybridoma cell strains of the positive clone secreting CDV-McAb were obtained.CE3 McAb against N protein was selected to prepare the FA.The FA was proved to be of highly specificity and sensitivity and can be used to detect CDV antigen for prevention and trcatment of CD.