Molecular detection of Peste des petits ruminant virus by using F, N and H genes based RT-PCR

Peste des petits ruminants; (PPR) is a severe viral disease of goats and sheep with high morbidity and sometimes high mortality characterized by fever, erosive stomatitis, conjunctivitis, gastroenteritis and pneumonia. PPR is caused by Peste des petits ruminants virus (PPRV), a Paramyxovirus of the Morbillivirus genus. The study involved the isolation of PPR virus from three clinical samples and assessment of F, N and H gene targets for detection of PPRV by RT-PCR from the clinical and cell culture samples. The isolation of PPRV in cell culture was confirmed after P 3, P 2 and P 2 passage by F, N and H gene based RT-PCR. A total of 23 samples including 15 cell culture samples, six tissue samples, one whole blood and reference vaccine virus (Sungri isolate) were processed for RNA extraction using TRI Reagent. RNA samples showing acceptable purity and concentration were reverse transcribed with random hexamers to generate cDNA templates, which were subjected to PCR amplification using F, N and H gene based primers. Reference vaccine virus as well as 10, 11 and 8 out of 15 cell culture samples of the three isolates produced approximately 372 bp amplicon with F1 / F2, 351 bp amplicon with NP3 / NP4 and 713 bp amplicon with H1 / H2 primers respectively.