Abstract
Potato is important in Western Australia both for domestic food production and export. Four viruses diminish tuber yield locally, Potato leaf roll virus (PLRV), Potato virus X (PVX), Potato virus S (PVS) and Tomato spotted wilt virus (TSWV). A real-time multiplex, single tube RT-PCR assay for the detection of these viruses from potato leaves and tubers was developed using Cy5-, FAM-, JOE- and ROX-labelled TaqMan probes. The copy numbers for transcripts were quantified with a dynamic range of 8x101 to 8x109 copies of PVX and PVS, 1x102 to 1x1010 copies of PLRV and 1x103 to 1x1010 copies of TSWV. The inter-assay reproducibility was high, with a coefficient of variation (CV) of <2%. Total RNA was rapidly and efficiently extracted from bulked tuber samples for the reliable detection of one or more of the viruses. These data indicate that this high-throughput test is accurate and sensitive, and will provide a cost-effective diagnostic tool for the seed potato industry.