Preparation of purified peste des petits ruminants (PPR) virus antigen from a local isolate of Bangladesh

Peste des petits ruminants (PPR) is the number one killer disease of small ruminants. The disease causes severe losses to small ruminant production and is presently considered as one of the major threats to about 22 million small ruminant population of Bangladesh where mortality may reach up to 100% in an outbreak. To control and eradicate PPR disease from Bangladesh quick detection and diagnosis is necessary. In field condition, PPR can be diagnosed by serological test and further confirmed by RT-PCR in laboratory. But all the tests are too expensive. The objective of the present study was to prepare purified Peste des petits ruminants virus (PPRV) antigen which can be used for the development of a quick and cheapest diagnostic ELISA kit. A recombinant tissue culture adapted strain of PPRV was propagated in Vero cell culture, the virus was pelleted by high speed centrifugation and the concentrated virus was purified by density gradient centrifugation using a discontinuous gradient of 30% and 60% sucrose. For further confirmation the purified virus was subjected to RT-PCR. The present method of preparing purified antigen appeared to be quite efficient as a distinct clear band of purified virus was found at the interface of 30% and 60% sucrose following density gradient centrifugation. In RT-PCR, a fragment of the Fusion (F) gene of purified PPRV was amplified with the expected band size of 448 bp. The purified PPR viral antigen may be used as a potential diagnostic antigen or immunogen.