Development of a Multiplex Reverse Transcription-Polymerase Chain Reaction Assay for the Simultaneous Detection of Three Viruses in Leguminous Plants

Abstract

A multiplex reverse transcription-polymerase chain reaction (mRT-PCR) assay was developed for the detection of Clover yellow vein virus (ClYVV), Peanut mottle virus (PeMoV), and Tomato spotted wilt virus (TSWV), which were recently reported to infect soybean and azuki bean in Korea. Species-specific primer sets were designed for the detection of each virus, and their specificity and sensitivity were tested using mixed primer sets. From among the designed primer sets, two combinations were selected and further evaluated to estimate the detection limits of uniplex, duplex, and multiplex RT-PCR. The multiplex RT-PCR assay could be a useful tool for the field survey of plant viruses and the rapid detection of ClYVV, PeMoV, and TSWV in leguminous plants.

A survey of viruses and viroids in astringent persimmon (Diospyros kakiThunb.) and the development of a one-step multiplex reverse transcription-polymerase chain reaction assay for the identification of pathogens

“…Furthermore, multiplex RT-PCR (mRT-PCR) allows rapid and low-cost detection owing to its ability to simultaneously detect multiple target pathogens in a single reaction (Asano et al 2015;Yao et al 2014). Therefore, diagnostic methods based on mRT-PCR have been developed for the diagnosis of viral pathogens in various plants, such as beans (Park et al 2018a), cherries (Park et al 2018b), garlic (Nam et al 2015), and pears (Kim et al 2019).…”