title: Development of a Rapid Detection Method for Potato virus X by Reverse Transcription Loop-Mediated Isothermal Amplification.
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.
authors.affiliation: Department of Agricultural Biology, College of Agriculture & Life Sciences, Chonbuk National University, Jeonju-si 561-756, Korea.
authors.forename: Joojin
authors.initials: J
authors.lastname: Jeong
authors.affiliation: Department of Agricultural Biology, College of Agriculture & Life Sciences, Chonbuk National University, Jeonju-si 561-756, Korea.
authors.forename: Sang-Yun
authors.initials: SY
authors.lastname: Cho
authors.affiliation: Department of Agricultural Biology, College of Agriculture & Life Sciences, Chonbuk National University, Jeonju-si 561-756, Korea.
authors.forename: Wang-Hyu
authors.initials: WH
authors.lastname: Lee
authors.affiliation: Division of Biotechnology, Chonbuk National University, Iksan 570-752, Korea ; Plant Medicinal Research Center, College of Agriculture & Life Sciences, Chonbuk National University, Jeonju-si 561-756, Korea.
authors.forename: Kui-Jae
authors.initials: KJ
authors.lastname: Lee
authors.affiliation: Department of Agricultural Biology, College of Agriculture & Life Sciences, Chonbuk National University, Jeonju-si 561-756, Korea ; Plant Medicinal Research Center, College of Agriculture & Life Sciences, Chonbuk National University, Jeonju-si 561-756, Korea.
authors.forename: Ho-Jong
authors.initials: HJ
authors.lastname: Ju
keywords: Potato virus X; RT-LAMP reaction; loop primer sets; virus detection