The Plant Pathology Journal

The Korean Society of Plant Pathology (한국식물병리학회)
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Development of a Rapid Detection Method for Potato virus X by Reverse Transcription Loop-Mediated Isothermal Amplification

  • Jeong, Joojin (Department of Agricultural Biology, College of Agriculture & Life Sciences, Chonbuk National University) ;
  • Cho, Sang-Yun (Department of Agricultural Biology, College of Agriculture & Life Sciences, Chonbuk National University) ;
  • Lee, Wang-Hyu (Department of Agricultural Biology, College of Agriculture & Life Sciences, Chonbuk National University) ;
  • Lee, Kui-jae (Division of Biotechnology, Chonbuk National University) ;
  • Ju, Ho-Jong (Department of Agricultural Biology, College of Agriculture & Life Sciences, Chonbuk National University)
  • Received : 2015.03.30
  • Accepted : 2015.05.10
  • Published : 2015.09.01
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Abstract

The primary step for efficient control of viral diseases is the development of simple, rapid, and sensitive virus detection. Reverse transcription loop-mediated isothermal amplification (RT-LAMP) has been used to detect viral RNA molecules because of its simplicity and high sensitivity for a number of viruses. RT-LAMP for the detection of Potato virus X (PVX) was developed and compared with conventional reverse transcription polymerase chain reaction (RT-PCR) to demonstrate its advantages over RT-PCR. RT-LAMP reactions were conducted with or without a set of loop primers since one out of six primers showed PVX specificity. Based on real-time monitoring, RT-LAMP detected PVX around 30 min, compared to 120 min for RT-PCR. By adding a fluorescent reagent during the reaction, the extra step of visualization by gel electrophoresis was not necessary. RT-LAMP was conducted using simple inexpensive instruments and a regular incubator to evaluate whether RNA could be amplified at a constant temperature instead of using an expensive thermal cycler. This study shows the potential of RT-LAMP for the diagnosis of viral diseases and PVX epidemiology because of its simplicity and rapidness compared to RT-PCR.

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References

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