The production and evaluation of a standard reference peste des petits ruminants (PPR)-diagnostic antigen using a sonication technique.

Abstract

Peste des petits ruminants (PPR) is a widely spread viral disease with limited sporadic outbreaks particularly among small ruminants. The diagnosis depends on the detection of antibodies by commercially available expensive reagents. The aim of this work was to prepare and evaluate antigen from an attenuated local PPR virus in Egypt. The virus was propagated in Vero cells, subjected to 3 cycles freeze/thaw, concentrated by dialysis and sonicated for 60 seconds at 3.5 power. The antigen was tested for sterility by re-passage several dilutions in Vero cells, and culture for aerobic/anaerobic bacteria, mycoplasma and fungi using selective media. The efficiency of the antigen was evaluated against reference antigen for reactivity using ELISA and agar gel immunodiffusion (AGID) tests. The prepared antigen proved to be completely inactivated, not contaminated and efficient for the detection of antibodies to PPR by both ELISA and AGID tests. In conclusion, the prepared antigen is reliable for serodiagnosis of PPR and can replace the expensive commercial antigenss. Local large scale production of this antigen will facilitate surveillance of the disease in Egypt.