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Development of RT-PCR and Nested PCR for Detecting Four Quarantine Plant Viruses Belonging to Nepovirus
Res. Plant Dis. 2013;19:220-225
Published online September 30, 2013
© 2013 The Korean Society of Plant Pathology.

Siwon Lee1,2, Eun-Ha Kang1,3, Yong-Gil Shin1* and Su-Heon Lee4,5**

1Plant Quarantine Technology Center, Animal and Plant Qurantine Agency, Suwon 443-440, Korea
2Department of Microbiology, Dankook University, Cheonan 330-714, Korea
3Department of Genetic Engineering, Sungkyunkwan University, Suwon 440-746, Korea
4School of Applied Biosciences, Kyungpook National University, Daegu 702-701, Korea
5Institute of Plant Medicine, Kyungpook National University, Daegu 702-701, Korea
*Corresponding author
Phone) +82-31-204-0918, Fax) +82-31-204-0668 E-mail) syg1286@korea.kr
**Corresponding author
Phone) +82-53-950-5763, Fax) +82-53-950-6758 E-mail) suheon@knu.ac.kr
Received March 25, 2013; Revised September 13, 2013; Accepted September 17, 2013.
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/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract
본 연구에서는 식물검역바이러스 4종(TBRV, ArMV, CLRV 및 GFLV)을 RT-PCR과 nested PCR 방법으로 진단 할 수 있는 방법을 개발하였다. 본 연구에서 개발한 방법은 모두 같은 PCR 조건으로 검사자에게 편리성과 신속성을 높여줄 뿐 아니라, 돌연변이-양성대조구의 사용으로 실험 오염여부를 확인할 수 있어 더욱 정확하다. 개발한 방법으로 최근 3년 Nepovirus속 4종의 바이러스를 검사한 결과, 27건을 검출하여 검역처분 하였다. 본 연구 결과들은 앞으로도 수출입 식물에서 해당 바이러스들을 신속, 정밀하게 진단할 수 있는 방법으로 활용할 수 있을 것으로 기대된다.
For quarantine purpose, we developed the RT- and nested PCR module of Tomato black ring virus (TBRV), Arabis mosaic virus (ArMV), Cherry leafroll virus (CLRV) and Grapevine fanleaf virus (GFLV). The PCR modules, developed in this study make diagnosis more convenient and speedy because of same PCR condition. And also, the methods are more accurate because it can check whether the result is contamination or not using the mutation-positive control. We discard or return the 27 cases of Nepovirus infection seed by employing the module past 3 years. This study provides a rapid and useful method for detection of four quarantine plant viruses.
Keywords : Detection, Nepovirus, Nested PCR, Quarantine, RT-PCR


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