Journal article
A single tube, quantitative real-time RT-PCR assay that detects four potato viruses simultaneously
Journal of Virological Methods, Vol.161(2), pp.289-296
2009
Abstract
A high throughput, real-time multiplex, single tube RT-PCR assay was developed for simultaneous detection of Potato leafroll virus (PLRV), Potato virus X (PVX) and Potato virus S (PVS) in potato leaves and tubers, and Tomato spotted wilt virus (TSWV) in potato tubers and tomato leaves. The test uses four different fluorescently labelled TaqMan® probes. Limits of detection sensitivity were established using a range of virus transcript copy numbers (8 × 101 to 8 × 109 copies of PVX and PVS, 1 × 102 to 1 × 1010 copies of PLRV and 1 × 103 to 1 × 1010 copies of TSWV). For each individual assay, the inter-assay reproducibility was high, with a coefficient of variation of the combined assays of <2%. Total RNA was extracted rapidly and efficiently from bulked samples equivalent to 300 dormant tubers to detect single infections of PLRV, PVX, PVS and TSWV simultaneously in a single assay. The multiplexed assay was validated in blind studies with leaves and tubers. This high-throughput test is accurate and sensitive, and provides seed potato industries with a cost-effective diagnostic tool to detect viruses reliably in bulked samples of dormant potato tubers.
Details
- Title
- A single tube, quantitative real-time RT-PCR assay that detects four potato viruses simultaneously
- Authors/Creators
- S.M. Mortimer-Jones (Author/Creator) - Murdoch UniversityM.G.K. Jones (Author/Creator) - Murdoch UniversityR.A.C. Jones (Author/Creator) - Murdoch UniversityG. Thomson (Author/Creator) - Murdoch UniversityG.I. Dwyer (Author/Creator) - Murdoch University
- Publication Details
- Journal of Virological Methods, Vol.161(2), pp.289-296
- Publisher
- Elsevier BV
- Identifiers
- 991005541548807891
- Copyright
- © 2009 Elsevier B.V.
- Murdoch Affiliation
- School of Biological Sciences and Biotechnology
- Language
- English
- Resource Type
- Journal article
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- Citation topics
- 3 Agriculture, Environment & Ecology
- 3.267 Virology - Plant
- 3.267.356 Plant Virus Interactions
- Web Of Science research areas
- Biochemical Research Methods
- Biotechnology & Applied Microbiology
- Virology
- ESI research areas
- Microbiology