Please use this identifier to cite or link to this item: https://hdl.handle.net/1959.11/3587
Title: Differential amplification and quantitation of Marek's disease viruses using real-time polymerase chain reaction
Contributor(s): Islam, Aminul (author); Harrison, Bruce (author); Cheetham, Brian Frederick  (author); Mahoney, Timothy J (author); Young, Peter L (author); Walkden-Brown, Steve William  (author)orcid 
Publication Date: 2004
DOI: 10.1016/j.jviromet.2004.03.006
Handle Link: https://hdl.handle.net/1959.11/3587
Abstract: Quantitative real-time PCR (qPCR) assays for the three serotypes of Marek's disease virus (MDV) have been developed. An internal control qPCR assay that detects chicken α2 (VI) collagen gene was also developed to allow quantitation of MDV. To reduce costs and time, the assays for MDV1 and the internal control were combined into a duplex assay. The sensitivity, specificity, precision, and reproducibility of each assay are reported. The MDV qPCR assays were specific to their target gene when compared using Australian field and vaccine strains of MDV and 10–100-fold more sensitive than standard PCR. Using DNA from infected spleen tissue, the lower limit of detection of total DNA (viral and host combined) was 0.025 ng for the MDV1 and collagen assays, and 0.25 ng for the HVT and MDV2 assays. All assays were found to be highly reproducible for Ct values, but less so for calculated concentrations. MDV1 and HVT were quantitated in spleen tissue of twenty experimentally infected chickens 7–35 days after infection. The relative abundance of MDV1 exhibited a clear peak at day 14 post-infection, whereas HVT displayed an increasing trend over the 35 days post-infection. The duplex assay was optimized such that it was able to accurately quantitate MDV1 in samples of very high, medium, and very low relative abundance of MDV1. These qPCR assays will be useful for reliable differentiation and quantitation of MDV for a range of research and industry applications.
Publication Type: Journal Article
Source of Publication: Journal of Virological Methods, 119(0), p. 103-113
Publisher: Elsevier BV
Place of Publication: Netherlands
ISSN: 1879-0984
0166-0934
Fields of Research (FoR) 2008: 060506 Virology
Socio-Economic Objective (SEO) 2008: 830309 Poultry
Peer Reviewed: Yes
HERDC Category Description: C1 Refereed Article in a Scholarly Journal
Appears in Collections:Journal Article

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