Abstract
Multiplex primers were calculated using a web-based program, muPlex in order to detect eight cucurbit-infecting viruses: Watermelon mosaic virus (WMV), Zucchini yellow mosaic virus (ZYMV), Cucumber mosaic virus (CMV), Kyuri green mottle mosaic virus (KGMMV), Papaya ringspot virus (PRSV), Melon yellow spot virus (MYSV), Cucumber green mottle mosaic virus (CGMMV), and Beet pseudo-yellows virus (BPYV). The predicted length of the amplified fragments ranged from 283 by to 1, 144 bp, and were distinct from each other. RT-PCR conducted using a mixture of all the primer pairs (primer cocktail) amplified the appropriate fragments corresponding to each virus species. Non-specific fragments were not observed. Two fragments were simultaneously amplified when RNA mixtures containing two distinct viral species were used as templates. However, if the concentration of one viral RNA was too high, the other virus could not be detected.
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