Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
6132740 | Journal of Virological Methods | 2016 | 24 Pages |
Abstract
For a simple and rapid detection of Chrysanthemum stem necrosis virus (CSNV) from chrysanthemum and tomato, a reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed. A primer set designed to the genome sequences of CSNV worked most efficiently at 63 °C and could detect CSNV RNA within 12 min by fluorescence monitoring using an isothermal DNA amplification and fluorescence detection device. The result of a specificity test using seven other viruses and one viroid-infectable chrysanthemum or tomato showed that the assay could amplify CSNV specifically, and a sensitivity comparison showed that the RT-LAMP assay was as sensitive as the reverse transcriptase polymerase chain reaction. The RT-LAMP assay using crude RNA, extracted simply, could detect CSNV. Overall, the RT-LAMP assay was found to be a simple, specific, convenient, and time-saving method for CSNV detection.
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Authors
Ryoji Suzuki, Shiro Fukuta, Yuho Matsumoto, Toru Hasegawa, Hiroko Kojima, Makiko Hotta, Noriyuki Miyake,