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- Title
Molecular diagnosis and adaptation of highly virulent infectious bursal disease virus on chicken embryo fibroblast cell.
- Authors
Singh, Yogender; Yadav, Kalpana; Singh, Vinod Kumar; Kumar, Rakesh
- Abstract
Aim: To collect Infectious bursal disease virus (IBDV) infected bursae based on postmortem findings of clinical samples followed by molecular confirmation and adaptation on chicken embryo fibroblast cell. Material and Methods: Enlarged bursae were processed to make 10% suspension in phosphate buffer saline and were used for viral RNA isolation to carryout VP2 gene fragment amplification using RT-PCR technique. Suspension was also used for adaptation of IBDVon chicken embryo fibroblast cells prepared from 11 day old chicken embryos. Infective titer of virus was calculated using Reed and Muench method. Results: Fifteen dead birds suspected of IBD infection were collected from different local poultry farms of Jabalpur (Madhya Pradesh). After post-mortem, twelve enlarged bursae sample were used for total RNA isolation which was used for amplification of VP2 gene. Outoftwelve, eleven were found positive for VP2 gene amplification. Virological characterization of PCR positive sample was done on chicken embryo fibroblast cell culture and various cytopathic effects like rounding of cells, cellular detachment and vacuolation were shown by five IBD field isolates after 48 hours on 4th passage. TCID50 perml of the adapted viruson 4th passage at 48 hours after infection was 1.46 x 107. Conclusion: VP2 gene amplification using RT-PCR technique is a specific target for IBDV detection. Passaging of highly virulent IBDV field isolatesin cell culture leads to attenuation of virus which can be exploited as a cell culture adapted vaccine candidate.
- Subjects
MOLECULAR diagnosis; INFECTIOUS bursal disease virus; CHICKEN embryos; FIBROBLASTS; AUTOPSY; NUCLEIC acid isolation methods
- Publication
Veterinary World, 2014, Vol 7, Issue 5, p351
- ISSN
0972-8988
- Publication type
Article
- DOI
10.14202/vetworld.2014.351-355