Development of reverse transcription loop-mediated isothermal amplification assay as a simple detection method of Chrysanthemum stem necrosis virus in chrysanthemum and tomato
Development of reverse transcription loop-mediated isothermal amplification assay as a simple detection method of Chrysanthemum stem necrosis virus in chrysanthemum and tomato
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DOI：
10.1016/j.jviromet.2016.07.005
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发表时间：
2016-10-01
影响因子：
3.1
通讯作者：
Miyake, Noriyuki
中科院分区：
文献类型：
--
作者：
Suzuki, Ryoji;Fukuta, Shiro;Miyake, Noriyuki
为了简便、快速地检测菊花和番茄中的菊花茎坏死病毒（Chrysanthemum stem necrosis virus，CSNV），建立了一种检测菊花和番茄中CSNV的逆转录环介导等温扩增（RT-LAMP）方法。针对CSNV基因组序列设计的引物组在63 ℃下最有效，并且可以使用等温DNA扩增和荧光检测装置通过荧光监测在12分钟内检测CSNV RNA。用其他7种病毒和1种类病毒感染的菊花或番茄进行的特异性试验结果表明，该检测方法能特异性扩增CSNV，灵敏度比较表明RT-LAMP检测方法与逆转录聚合酶链反应一样灵敏。用简单提取的粗RNA进行RT-LAMP检测，可以检测到CSNV。总的来说，RT-LAMP检测被认为是一种简单、特异、方便和节省时间的CSNV检测方法。(C)2016爱思唯尔B. V.保留所有权利。
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 degrees 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. (C) 2016 Elsevier B.V. All rights reserved.