Isolation of infectious bursal disease virus from cell culture and bursal tissue and confirmation by RT-PCR and electron microscopy
S Upadhyay, V Umapathi and BK Sarkar
In this study, isolation of the infectious bursal disease virus was carried out from two different sources viz. virus adapted in cell culture and bursal tissue obtained from field outbreaks. The cell culture adapted UA/BZ/2 strain of the virus was propagated in chicken embryo fibroblast upto 8th passage and 1000 ml of the viral suspension was prepared. The bursal tissue from infected chickens obtained from the field outbreak cases of Infectious Bursal Disease in Hisar, Haryana was triturated and 10% suspension in phosphate buffered saline was prepared. Both the suspensions were then subjected to centrifugatiion and ultracentrifugation to obtain the partially purified virus. The presence of the virus in the final suspension was confirmed by AGPT and RT-PCR by amplifying 248 bp fragment of the VP2 hypervariable region of the virus. Further electron microscopy confirmed the IBD viruses as classical birna group of viruses on the basis of morphological structure.