Research in Plant Disease (식물병연구)

The Korean Society of Plant Pathology (한국식물병리학회)
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Reverse Transcription Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Soybean yellow mottle mosaic virus

콩황화모틀모자이크바이러스의 신속검출을 위한 역전사 등온증폭법

  • Bae, Dae Hyeon (School of Applied Biosciences, Kyungpook National University) ;
  • Park, Chung Youl (School of Applied Biosciences, Kyungpook National University) ;
  • Kim, Bong-Sub (School of Applied Biosciences, Kyungpook National University) ;
  • Lee, Yeong-Hoon (Crop Production Technology Research Division, Department of Southern Area Crop Science, National Institute of Crop Science, Rural Development Administration) ;
  • Yoon, Young-Nam (Crop Production Technology Research Division, Department of Southern Area Crop Science, National Institute of Crop Science, Rural Development Administration) ;
  • Kang, Hang Won (Crop Production Technology Research Division, Department of Southern Area Crop Science, National Institute of Crop Science, Rural Development Administration) ;
  • Oh, Jonghee (School of Applied Biosciences, Kyungpook National University) ;
  • Lee, Su-Heon (School of Applied Biosciences, Kyungpook National University)
  • 배대현 (경북대학교 응용생명과학부) ;
  • 박충열 (경북대학교 응용생명과학부) ;
  • 김봉섭 (경북대학교 응용생명과학부) ;
  • 이영훈 (농촌진흥청 국립식량과학원 남부작물부 생산기술개발과) ;
  • 윤영남 (농촌진흥청 국립식량과학원 남부작물부 생산기술개발과) ;
  • 강항원 (농촌진흥청 국립식량과학원 남부작물부 생산기술개발과) ;
  • 오종희 (경북대학교 응용생명과학부) ;
  • 이수헌 (경북대학교 응용생명과학부)
  • Received : 2016.06.01
  • Accepted : 2016.08.11
  • Published : 2016.09.30
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Abstract

Soybean yellow mottle mosaic virus (SYMMV) is a new emerging plant virus detected in soybean (Glycine max) in Korea. Reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for rapid detection of SYMMV has been developed. In this study, we have designed primers (SYMM-F3/B3/FIP/BIP) specific to sequences from the coat protein gene of SYMMV genome. Sensitivity analysis showed that RT-LAMP was 10 to 100 times more sensitive than reverse transcription polymerase chain reaction (RT-PCR). The optimal reaction condition of RT-LAMP was determined at $65^{\circ}C$ for 50 minutes. The result indicates that RT-LAMP assay does not require special equipment and long time for SYMMV detection. Therefore, it can be an alternative detection method of RT-PCR in laboratory.

SYMMV는 콩에서 빈번하게 발생하는 바이러스이며, 이 바이러스의 발생률은 계속해서 증가하고 있다. 본 연구에서는 콩에 발생하는 SYMMV를 신속하게 검출하기 위해서 RT-LAMP를 적용하였다. RT-LAMP 방법은 등온에서 단시간에 유전자 증폭이 가능하고, 전기영동 없이도 형광물질을 이용해 바이러스를 검출할 수 있는 이점이 있다. 프라이머는 SYMMV coat protein gene의 염기서열을 기반으로 4개의 프라이머를 설계하였다. 실험결과 SYMMV RT-LAMP는 $65^{\circ}C$에서 50분간 증폭시켰을 때 최적의 효율을 보였다. 또한, RT-LAMP와 RT-PCR과의 민감도를 비교한 결과 RT-LAMP 방법이 10-100배 정도 더 우수한 민감도를 가지는 것으로 밝혀졌다. 본 실험 결과를 토대로 기존의 진단법과 비교하여 높은 민감도와 짧은 소요시간에 이점이 있는 RT-LAMP는 SYMMV의 현장 및 연구실에서의 진단에 적용될 수 있을 것이라 생각된다.

Keywords

References

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