One-Step Multiplex RT-PCR Method for Detection of Melon Viruses

This study presents a one-step multiplex reverse transcription polymerase chain reaction (RT-PCR) method for the simultaneous detection of multiple viruses affecting melon crops. Viruses such as <i>Watermelon mosaic virus</i> (WMV), <i>Cucumber mosaic virus</i> (CMV), <i>Zucchini yellow mosaic virus</i> (ZYMV), <i>Squash mosaic virus</i> (SqMV), <i>Tobacco mosaic virus</i> (TMV), <i>Papaya ring spot virus</i> (PRSV), and <i>Melon yellow spot virus</i> (MYSV) pose a great threat to melons. The mixed infection of these viruses is the most common observation in the melon-growing fields. In this study, we surveyed northern Xingjiang (Altay, Changji, Wujiaqu, Urumqi, Turpan, and Hami) and southern Xingjiang (Aksu, Bayingolin, Kashgar, and Hotan) locations in Xinjiang province and developed a one-step multiplex RT-PCR to detect these melon viruses. The detection limits of this multiplex PCR were 10³ copies/μL for ZYMV and MYSV and 10² copies/μL for WMV, SqMV, PRSV, CMV, and TMV. The detection results in the field showed 242 samples were infected by one or more viruses. The multiplex RT-PCR protocol demonstrated rapid, simultaneous, and relatively effective detection of viruses such as WMV, CMV, ZYMV, SqMV, TMV, PRSV, and MYSV. The technique is designed to identify these melon viruses in a single reaction, enhancing diagnostic efficiency and reducing costs, thus serving as a reference for muskmelon anti-virus breeding in Xinjiang.