Absolute quantification using real-time polymerase chain reaction of Marek's disease virus serotype 2 in field dust samples, feather tips and spleens

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 disea...

Full description

Saved in:
Bibliographic Details
Published in:Journal of virological methods 2006-08, Vol.135 (2), p.186-191
Main Authors: Renz, Katrin G., Islam, Aminul, Cheetham, Brian F., Walkden-Brown, Stephen W.
Format: Article
Language:English
Subjects:
Absolute quantification
Animals
Biological and medical sciences
Chickens - virology
Dust
Feathers - virology
Fundamental and applied biological sciences. Psychology
Herpesvirus 3, Gallid - classification
Herpesvirus 3, Gallid - genetics
Herpesvirus 3, Gallid - immunology
Herpesvirus 3, Gallid - isolation & purification
Marek's disease herpesvirus
Marek's disease virus serotype 2
Microbiology
Plasmids
Polymerase Chain Reaction - methods
qPCR
Real-time PCR
Reproducibility of Results
Sensitivity and Specificity
Spleen - virology
Techniques used in virology
Vaccination
Viral Vaccines - immunology
Virology
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary: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 intra- and 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.
ISSN:0166-0934
1879-0984
DOI:10.1016/j.jviromet.2006.03.017