Journal
JOURNAL OF VIROLOGICAL METHODS
Volume 135, Issue 2, Pages 186-191Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jviromet.2006.03.017
Keywords
real-time PCR; qPCR; absolute quantification; Marek's disease virus serotype 2
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Methods for Taqman quantitative real-time PCR (qPCR) assays to detect the three serotypes of Marek's disease virus (MDV) are available, and absolute quantification has been developed for MDV serotype 1 and serotype 3. The development of a method for absolute quantification of Marek's disease virus serotype 2 (MDV2) is described in this paper. Using plasmid DNA, the lower detection limit of the MDV2 assay was determined to be 10 copies. Three independent assay runs showed highly reproducible Ct values and calculated copy numbers, with mean intraand inter-assay coefficients of variation of less than 3% for Ct and less than 21.5% for calculated copy number. Absolute quantification of MDV2 was performed successfully on dust samples collected from poultry farms across Australia, material from infectious spleens and feather tips from chickens vaccinated with an attenuated strain of MDV2. Thus, it is now possible to use qPCR assays for absolute quantification of all three serotypes of MDV in a sample. (c) 2006 Elsevier B.V. All rights reserved.
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