Isolation of infectious bursal disease virus in cell culture, detection and characterization by RT-PCRSSCP analysis

Gaba, Amit; Makadiya, N.R.; Joshi, C.G. and Jhala, M.K. (2007) Isolation of infectious bursal disease virus in cell culture, detection and characterization by RT-PCRSSCP analysis. Indian Journal of Poultry Science, 3 (42). pp. 339-342.

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Abstract

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 cwell monolayers inoculated with two field bursal samples resulted in isolation of the virus after five succesive passages. Distinct CPE charcaterized by rounding and clumping of cells, extensive cytolasmic granularity and detachmenet of the cells was obsorved, which was similar to that produced by vaccine isolated 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 isoalted and vaccine virus revealed similar migration pattern, suggesteing absence of any nucleotide sequence variation among them with respect to the targated gene segment.

EPrint Type:Article
Additional Information:Gumboroo - Infectious viral disease - IBD - IBDV
Uncontrolled Keywords:RT-PCR, BGM-70, Cell culture, IBDV
Subjects:Animal Structures > Bursa of Fabricius
-Institutional Repositories > College of Veterinary Science and Animal Husbandry (Anand Agricultural University)
Genetic Structures > Genes
Animal Diseases > Bird Diseases > Poultry Diseases
Biological Sciences > Biotechnology
Virus Diseases > RNA Virus Infections > Birnaviridae Infections
ID Code:2975
Deposited By:Dr Prakash Koringa
Deposited On:31 July 2008

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