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

Islam, Aminul, Cheetham, Brian F., Mahony, Timothy J., Young, Peter L. and Walkden-Brown, Stephen W. (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: 127-134.


Author Islam, Aminul
Cheetham, Brian F.
Mahony, Timothy J.
Young, Peter L.
Walkden-Brown, Stephen W.
Title Absolute quantitation of Marek's disease virus and Herpesvirus of turkeys in chicken lymphocyte, feather tip and dust samples using real-time PCR
Journal name Journal of Virological Methods  (ERA 2012 Listed)    (ERA 2010 Rank B)   Check publisher's open access policy
Publication date 2006-03
Sub-type Article
DOI 10.1016/j.jviromet.2005.10.009
Volume number 132
Issue number 1-2
ISSN 0166-0934; 1879-0984
Start page 127
End page 134
Total pages 8
Place of publication Amsterdam, Netherlands
Publisher Elsevier BV
Language eng
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.
Keyword Real-time Pcr
Qpcr
Viral Quantitation
Marek's Disease Virus Serotype 1
Herpesvirus Of Turkeys
Polymerase-chain-reaction
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Non-UQ

Document type: Journal Article
Sub-type: Article
Collection: Queensland Alliance for Agriculture and Food Innovation
 
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Created: Mon, 07 Mar 2011, 15:17:30 EST