title: Isolation of bovine viral diarrhea virus 1, a pestivirus from autopsied lamb specimen from Tamil Nadu, India.
abstract: An epizootic of febrile illness among the Madras red breed of sheep had occurred in 1994 in Verrapuram, Chennai, India. The epizootic was suspected as Rift Valley fever (RVF)-like sickness based on clinical features. However, its etiological agent could neither be isolated nor implicated conclusively. During the post-epizootic period a male lamb died of similar clinical features and the spleen was immediately collected. Inoculation of spleen suspension in infant mouse brain yielded a virus that was serially passaged in infant mice and rhabdomyosarcoma (RD) cells. Electron microscopic observations revealed virus particles resembling flaviviruses. RT-PCR performed on extracted total RNA from infected cells and mouse brains with flavivirus-specific or RVF-specific primers gave negative results. However, an amplicon of 280 bp was obtained with pestivirus-specific primers from the 5'-UTR. Further, a nested PCR yielded a product of 157 bp. Nucleotide sequencing of the 157 bp product showed 100% homology to BVDV-1. Western blot analysis with a flavivirus envelope protein-specific MAb revealed three proteins of 33 K, 45 K and 55 K. Further studies suggested that the 33 K and 55 K proteins were glycosylated. This is the first report of isolation of BVDV-1 from a lamb in India.
authors.affiliation: National Institute of Virology, 20-A, Dr. Ambedkar Road, Pune 411 001, India.
authors.forename: P
authors.initials: P
authors.lastname: Yadav
authors.forename: P V
authors.initials: PV
authors.lastname: Barde
authors.forename: R
authors.initials: R
authors.lastname: Jadi
authors.forename: M D
authors.initials: MD
authors.lastname: Gokhale
authors.forename: A
authors.initials: A
authors.lastname: Basu
authors.forename: M V
authors.initials: MV
authors.lastname: Joshi
authors.forename: R
authors.initials: R
authors.lastname: Mehla
authors.forename: S R P
authors.initials: SR
authors.lastname: Kumar
authors.forename: S S
authors.initials: SS
authors.lastname: Athavale
authors.forename: D T
authors.initials: DT
authors.lastname: Mourya
mesh_terms: D000818:Animals; D001344:Autopsy; D015153:Blotting, Western; D030241:Diarrhea Virus 1, Bovine Viral / Q000235:genetics / Q000302:isolation & purification*; D006031:Glycosylation; D008297:Male; D051379:Mice; D008854:Microscopy, Electron; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012756:Sheep / Q000821:virology*