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Absolute quantitation of Marek's disease virus and Herpesvirus of turkeys in chicken lymphocyte, feather tip and dust samples using real-time PCR

Islam, A. and Walkden-Brown, S.W. and Cheetham, B.F. and Mahony, T.J. and Young, P.L. (2006) Absolute quantitation of Marek's disease virus and Herpesvirus of turkeys in chicken lymphocyte, feather tip and dust samples using real-time PCR. Journal of Virological Methods, 132 (1-2). pp. 127-134.

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Article Link(s): http://dx.doi.org/10.1016/j.jviromet.2005.10.009

Publisher URL: http://www.elsevier.com

Abstract

The further development of Taqman quantitative real-time PCR (qPCR) assays for the absolute quantitation of Marek's disease virus serotype 1 (MDV1) and Herpesvirus of turkeys (HVT) viruses is described and the sensitivity and reproducibility of each assay reported. Using plasmid DNA copies, the lower limit of detection was determined to be 5 copies for the MDV1 assay and 75 copies for the HVT assay. Both assays were found to be highly reproducible for Ct values and calculated copy numbers with mean intra- and inter-assay coefficients of variation being less than 5% for Ct and 20% for calculated copy number. The genome copy number of MDV1 and HVT viruses was quantified in PBL and feather tips from experimentally infected chickens, and field poultry dust samples. Parallelism was demonstrated between the plasmid-based standard curves, and standard curves derived from infected spleen material containing both viral and host DNA, allowing the latter to be used for absolute quantification. These methods should prove useful for the reliable differentiation and absolute quantitation of MDV1 and HVT viruses in a wide range of samples.

Item Type:Article
Corporate Creators:Emerging Technologies
Additional Information:© Elsevier B.V.
Keywords:Herpesvirus of turkeys; Marek's disease virus serotype 1; qPCR; real-time PCR; viral quantitation.
Subjects:Veterinary medicine > Diseases of special classes of animals > Avian
Science > Biology > Genetics
Veterinary medicine > Veterinary virology
Deposited On:23 Feb 2009 03:34
Last Modified:09 Sep 2010 02:03

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