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Authors: Pawan Kumar
Advisor: Gaya Prasad
Title: Molecular serotyping of Indian isolates of bluetongue virus based on vp2 gene
Publisher: LUVAS
Language: en
Type: Thesis
Keywords: Bluetongue virus (BTV), Nucleic acid sequencing, RT-PCR, Serotyping, Vp2 gene
Abstract: Twenty-four distinct serotypes of Bluetongue virus (BTV) has been recognized to cause one of the most important vector borne viral disease of ruminants worldwide out of which 21 are reported from different geographical areas of India (Prasad, 2000). BTV typing based on most divergent vp2 gene is crucial for the development of vaccine to control the disease. Conventional antibody based serotyping is time consuming and may result in the erroneous conclusions due to cross reactivity of different serotypes. Therefore, the present study was carried out for molecular typing of new Indian BTV isolates, based on RT-PCR and nucleotide sequencing of hypervariable regions of vp2 gene. All the six suspected isolates were confirmed as BTV based on characteristic cytopathic effects in BHK-21 cell line, characteristic 3:3:3:1 migration pattern of dsRNA genome segments in RNA-PAGE and BTV group specific RT-PCR assay. BTV isolates were then typed using type specific RT-PCR and confirmed by nucleotide sequencing. KOL-1 (West Bengal) and IVRI-2 (U.P.) isolates were typed as BTV3, ONG-3 and ONG-5 (A.P) as BTV9, VJW-64 (A.P.) as BTV16 and KMNO-7 (A.P.) as BTV21. The serotype of isolates was confirmed by BLAST and multiple sequence alignment with the gene sequences available in GenBank. KOL-1 and IVRI-2 were 9-11 % divergent from other BTV3 isolates, ONG-3 and ONG-5 isolates were 3-4 % divergent from BTV9K8IND and BTV9ITA isolates, VJW-64 was 4-5 % divergent from other BTV16 isolates whereas KMNO-7 isolate was 5 % divergent from BTV21SA and BTV21AUS isolates. BTV3 Indian isolates (KOL-1 and IVRI-2) were 96% and BTV9 Indian (ONG-3 and ONG-5) isolates were 99 % homologous. No significant similarity was observed between isolates of different serotypes based on their partial vp2 gene sequence. The present study for the first time revealed the presence of BTV21 serotype in India. BTV3 and BTV16 isolates, although reported earlier by typing in world reference laboratories, but in India we have identified and characterized these isolates for the first time. The study has generated new information about prevalence of BTV serotypes and thus may have an implication in future for development of effective vaccine in India.
Subject: Animal Biotechnology
Theme: Molecular serotyping of indian isolates of bluetongue virus based on vp2 gene
These Type: M.V.Sc.
Issue Date: 2009
Appears in Collections:Thesis

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