Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
6134904 | Journal of Virological Methods | 2011 | 4 Pages |
Abstract
Both conventional and real time PCR (rt-PCR) assays based on the amplification of a 103Â bp fragment from the DNA polymerase (DNA pol) gene (conserved, non-structural) of Orf virus (ORFV) were developed for detection and semi-quantitation of ORFV DNA from infected cell culture and clinical samples. The latter technique was based on TaqMan chemistry. The rt-PCR assay was specific and sensitive as it could detect as low as 3.5Â fg or 15 copies of ORFV genomic DNA. Both intra- (0.38-1.0%) and inter-assay (0.53-2.87%) variabilities of rt-PCR were within the acceptable range meaning the high efficiency and reproducibility of the assay. The rt-PCR was applied successfully to detect ORFV DNA from suspected clinical samples. Further, the assay has shown a relative diagnostic sensitivity and specificity of 100% and 93.5%, respectively, when compared to B2L gene based semi-nested PCR implying a wide potential of this rt-PCR for rapid field diagnosis of Orf in sheep and goats.
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Authors
D.P. Bora, G. Venkatesan, V. Bhanuprakash, V. Balamurugan, M. Prabhu, M.S. Siva Sankar, R. Yogisharadhya,