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Comparative Polypeptide Profiling: Isolation, Propagation and Purification of Indian Isolates of Buffalopox Virus

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Author(s): Singh, R.K. | V. Balamurugan | M. Hosamani | C.C. Satheesh | T.J. Rasool | M.P. Yadav

Journal: Journal of Animal and Veterinary Advances
ISSN 1680-5593

Volume: 5;
Issue: 3;
Start page: 260;
Date: 2006;
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Keywords: Buffalopox virus | Indian isolates | isolation | propagation | purification

ABSTRACT
Buffalopox is a contagious viral disease-affecting buffaloes (Bubalus bubalis) and rarely cows, with morbidity up to 80%in affected herd causing high economic losses to the farmers. In the present study, as a preliminary work, the field isolates of BPV recovered from suspected clinical materials obtained from outbreaks of different geographical locations of the country were propagated in Vero cells and purified along with reference BP4 virus for comparison. Purification of BPV was carried out in sucrose density gradient (60-36%), where two white opalescent bands appeared after ultracentifugation. The upper band, above the 36% sucrose layer was found to be enveloped virus (Extra cellular enveloped virus) where as the band at interface was the pure naked virus (Intracellular Mature Virus). The purity of the virus preparations was assessed by UV-spectrophotometry, SDS-PAGE and infectivity assay in cell culture. The extinction ratio values (O.D. 260/280) for all the BPV isolates were within the range of 1.2 to 1.4 indicating highly purified nature of the BPV. On the basis of polypeptide compositions, BPV isolates were similar and contained more than 25 polypeptides with a molecular weight range from 14.2 kDa to 180 kDa. Both the opalescent bands produced characteristics CPE in Vero cell line in infectivity assay. This preliminary study will help in undertaking further work on protein profile and further characterization, which, in turn, will help in understanding the virological and immunological properties of the new virus isolates for development of suitable immuno-diagnostics and prophylactics.

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