Isolation and Molecular Characterization of Infectious Bovine Rhinotracheitis Virus.
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Date
2011-08-16
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Karnataka Veterinary, Animal and Fisheries Sciences University , Bidar
Abstract
Infectious Bovine Rhinotracheitis is one of the important viral disease of cattle
caused by Bovine herpes Virus -1. In India, IBR causes great economic loss to the
farmers and its prevalence is in increasing trend over the past 10 years. The present study
was undertaken to isolate and characterise the BoHV-1 isolates circulating in Karnataka
state. Samples such as Nasal swabs, conjunctival swabs and aborted foetal liver were
collected from 40 cattle showing symptoms similar to IBR. All the samples were
subjected for virus isolation in MDBK cell line. Among the 40 samples only three
samples yielded BoHV-1 isolates in cell culture which were confirmed by Virus
neutralisation test. A sensitive and specific PCR was developed targeting conserved
region of gC gene and applied directly to the DNA extracted from field samples. The
sensitivity of the designed gC specific primers was 75 per cent and compared with the
published primers targeting gB gene whose sensitivity was 66 per cent. The gC based
PCR products were subjected for restriction enzyme analysis which further confirmed the
isolates as BoHV-1. The amplified product of gC based PCR was cloned in pGEMT
vector and sequenced. The nucleotide sequences of gC gene of BoHV-1 field isolates viz.,
PD_ADMAS707/10, PD_ADMAS723/10, PD_ADMAS741/10 along with the reference
virus were compared with published sequences using the BLAST programme and
submitted to GenBank under the accession numbers JN022593, JN022594, JN171865
respectively. Phylogenetic analysis revealed that all three field isolates were clustered
with BoHV-1.1 subtype. Thus, the study revealed the persistence of BoHV-1.1 subtype in
India.
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P.G. Thesis
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