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Development of a Multiplex Reverse Transcription-Polymerase Chain Reaction Assay for the Simultaneous Detection of Three Viruses in Leguminous Plants
Res. Plant Dis. 2018;24:348-352
Published online December 31, 2018
© 2018 The Korean Society of Plant Pathology.

Chung Youl Park1,2, Hyun-Geun Min1, Hong-Kyu Lee1, Rameswor Maharjan3, Youngnam Yoon3*, and Su-Heon Lee1,4*

1School of Applied Biosciences, Kyungpook National University, Daegu 41566, Korea
2Present Address: Seed Testing & Research Center, Korea Seed & Variety Service, Gimcheon 39660, Korea
3Crop Production Technology Research Division, NICS, RDA, Miryang 50426, Korea
4Institute of Plant Medicine, Kyungpook National University, Daegu 41566, Korea
*Y. Yoon Tel: +82-55-350-1264 Fax: +82-55-352-3059 E-mail: yoonyn@korea.kr
S.-H. Lee Tel: +82-53-950-5763 Fax: +82-53-950-6758 E-mail: suheon@knu.ac.kr
Received September 17, 2018; Revised November 1, 2018; Accepted November 2, 2018.
cc This is an open access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
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.
Keywords : Clover yellow vein virus, Multiplex RT-PCR, Peanut mottle virus, Primer combinations, Tomato spotted wilt virus


December 2018, 24 (4)