Simultaneous detection of three Potyvirus species from Cucurbitaceae crops by multiplex RT-PCR.

A multiplex RT-PCR procedure was developed for the simultaneous detection of Watermelon mosaic virus (WMV), Zucchini yellow mosaic virus (ZYMV) and Papaya ringspot virus (PRSV) from Cucurbitaceae crops. Using oligo dT primer, cDNA was synthesized from virus RNA and amplified with a mixture of three primer sets specific to WMV, ZYMV and PRSV. Three potyviruses were accurately detected from cucumber plants singly, doubly or triply infected with these viruses as well as mixtures of virus RNA. With this method, three potyviruses could be detected from various kinds of naturally infected Cucurbitaceae crops more accurately than with indirect ELISA and more easily than with uniplex RT-PCR.