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Testing for potato viruses is globally very important to prevent a critical shortage of potato supply. In most countries, testing is obligated by law. In Germany, seed potatoes are monitored for six viruses: PLRV, PVY, PVM, PVA, PVX and PVS. They can cause up to 90% loss of potato tubers in the field. Common methods currently used for testing are ELISA and conventional real-time PCR, but both are very time-consuming, and the former needs a high capacity of green houses and human resources, the latter elaborate RNA extraction steps. Recently, we proposed a new method called real-time DiRT-PCR which enables us to test for PLRV, PVY and PVS along with an internal control in three duplex real-time PCR reactions directly on diluted tuber sap. In this study, we describe the first TaqMan® assay for PVM published so far and embed it into a multiplex system to detect the remaining viruses. We are now able to sensitively test for the presence of six viruses in two multiplex reactions using the real-time DiRT-PCR without RNA purification.

european journal of plant pathology

Development of the first PVM TaqMan® primer set and a one-step real-time multiplex DiRT-PCR for the detection of PLRV, PVY, PVM, PVS, PVA and PVX in potato tuber sap