Bluetongue disease is an economically important disease of small ruminants, which is transmitted by culicoides vectors. The disease is caused by an Orbivirus classified under Reoviridae family. The virus consists of ten segments of double stranded RNA as genetic material enclosed by the capsid proteins. The outer capsid consists of VP2 and VP5 proteins, among them the VP2 protein harbor predominant serotype specific epitopes. In the present study the field isolate of bluetongue virus was plaque purified for three times in vero cell lines. The ds RNA genome was extracted by acid phenol method revealed the characteristic double stranded RNA genome of the bluetongue virus. The cDNA was synthesized by reverse transcription and subjected for serotyping. Three sets of overlapping primers were designed based on the VP2 gene of BT virus serotype 10 reference strain. The RT-PCR assay was standardized for amplification of VP2 gene of plaque purified field isolate of BT virus. The results revealed that all three sets of primers were successfully amplified the VP2 gene with amplicon sizes of 905 bp, 806 bp and 811 bp for set 1, 2 and set 3 primers respectively. The designed primers specifically identified the field isolate of BT virus as serotype 10 and can be applied as a diagnostic tool for serotyping of BT virus isolates.

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