Microbiology and Biotechnology Letters (한국미생물·생명공학회지)

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Development of a Rapid Assay for Peach Rosette Mosaic Virus Using Loop-mediated Isothermal Amplification

Peach rosette mosaic virus 검출을 위한 신속한 등온증폭법 개발

  • Lee, Siwon (Environmental Infrastructure Research Department, National Institute of Environmental Research) ;
  • Lee, Jin-Young (Department of Microbiology, College of Natural Sciences, Dankook University) ;
  • Kim, Jin-Ho (Department of Chemistry, College of Natural Science, Dankook University) ;
  • Rho, Jae-Young (Department of Microbiology, College of Natural Sciences, Dankook University)
  • 이시원 (국립환경과학원 환경기반연구) ;
  • 이진영 (단국대학교 자연과학대학 미생물학과) ;
  • 김진호 (단국대학교 자연과학대학 화학과) ;
  • 노재영 (단국대학교 자연과학대학 미생물학과)
  • Received : 2016.09.12
  • Accepted : 2016.11.01
  • Published : 2016.12.28
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Abstract

Peach rosette mosaic virus (PRMV) is a plant virus that was first reported in 1933 by Peach. It can infect hosts including peach, grape, blueberry, dandelion, plum, cherry tree, and weeds. PRMV is non-reportable in Korea, but it is designated as a controlled virus requiring plant quarantine. In this study, for the rapid and specific detection of PRMV, we developed an assay using loop-mediated isothermal amplification (LAMP). Comparison between conventional polymerase chain reaction (PCR) methods (real time-PCR and nested PCR) and LAMP for the detection of PRMV revealed an equivalent level of sensitivity by all the tested methods. For the LAMP assay, outer primer sets were used to amplify a 264-bp PCR product, which was then digested using the restriction enzyme Pvu II (CAG/CTG), and the visualization of two digestion fragments (207 + 57 bp) indicated a positive reaction. The developed LAMP assay for PRMV is expected to enable the rapid monitoring of PRMV in plants.

Peach rosette mosaic virus (PRMV)는 1933년 복숭아에서 처음 보고되었으며, 복숭아, 자두, 블루베리, 민들레, 벚나무 등에 감염되는 식물바이러스이다. PRMV는 한국에서 보고된 적이 없으나, 식물검역에서 관리병(control viruses)으로 지정되어 있다. 이번 연구에서는 PRMV를 더욱 신속하고 특이적으로 진단하기 위하여 Loop-mediated isothermal amplification 분석법을 적용한 진단법을 개발하였다. LAMP 방법은 기존의 PCR 방법(RT-PCR 및 nested PCR)과 같은 검출 강도를 가지고 있다. 또한 LAMP 반응을 확인하기 위해 PRMV cDNA을 outer primer sets (Product size 264 bp)로 PCR 한 뒤, Pvu II (CAG/CTG) 제한효소를 처리하였다. 제한효소 처리 결과 2개의 digestion fragments (207 + 57 bp)가 확인되었다. PRMV의 LAMP 진단 방법은 관련 식물로부터 더욱 신속한 모니터링이 가능할 것으로 기대된다.

Keywords

References

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