Department of Veterinary Microbiology, College of Veterinary Science and Animal Husbandry, Anand Agricultural University, Anand - 388 001, (Gujarat).
*Corresponding author: E-mail: mkjhala_2003@yahoo.co.in
The study was aimed at isolation of infectious bursal disease virus (IBDV), detection by RT-PCR and genetic characterization by single strand conformational polymorphism (SSCP). BGM-70 cell monolayers inoculated with two field bursal samples resulted in isolation of the virus after five successive passages. Distinct CPE characterized by rounding and clumping of cells, extensive cytoplasmic granularity and detachment of the cells was observed, which was similar to that produced by vaccine isolate during first passage. RT-PCR performed for viral RNA extracted by Trizol Reagent from cell culture supernatant and infected monolayers, using P1 and P2 pair of primers specific to hypervariable region of VP2 gene, resulted in generation of a targeted amplicon of 643 bp, confirming the growth of IBDV in cell culture. On SSCP analysis, RT-PCR amplified products of both the field isolates and vaccine virus revealed similar migration pattern, suggesting absence of any nucleotide sequence variation among them with respect to the targeted gene segment.
IBD, IBDV, cell culture, BGM-70, RT-PCR, SSCP
