Relative quantitation goes viral: an RT-qPCR assay for a grapevine virus

Bester, R., Pepler, P.T. , Burger, J.T. and Maree, H.J. (2014) Relative quantitation goes viral: an RT-qPCR assay for a grapevine virus. Journal of Virological Methods, 210, pp. 67-75. (doi:10.1016/j.jviromet.2014.09.022) (PMID:25286180)

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Abstract

Accurate detection and quantitation of viruses can be beneficial to plant–virus interaction studies. In this study, three SYBR green real-time RT-PCR assays were developed to quantitate grapevine leafroll-associated virus 3 (GLRaV-3) in infected vines. Three genomic regions (ORF1a, coat protein and 3′UTR) were targeted to quantitate GLRaV-3 relative to three stably expressed reference genes (actin, GAPDH and α-tubulin). These assays were able to detect all known variant groups of GLRaV-3, including the divergent group VI, with equal efficiency. No link could be established between the concentration ratios of the different genomic regions and subgenomic RNA (sgRNA) expression. However, a significant lower virus concentration ratio for plants infected with variant group VI compared to variant group II was observed for the ORF1a, coat protein and the 3′UTR. Significant higher accumulation of the virus in the growth tip was also detected for both variant groups. The quantitation of viral genomic regions under different conditions can contribute to elucidating disease aetiology and enhance knowledge about virus ecology.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Pepler, Dr Theo
Authors: Bester, R., Pepler, P.T., Burger, J.T., and Maree, H.J.
College/School:College of Medical Veterinary and Life Sciences > Institute of Biodiversity Animal Health and Comparative Medicine
Journal Name:Journal of Virological Methods
Publisher:Elsevier B.V.
ISSN:0166-0934
ISSN (Online):1879-0984

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