大豆科学2012，Vol.31Issue(6)：980-983,987,5.
应用逆转录环介导等温扩增技术检测大豆种子中烟草环斑病毒
Detection of Tobacco Ringspot Virus in Soybean Seeds by the Reverse Transcription Loop-mediated Isothermal Amplification Method
摘要
Abstract
A sets of primers were designed and synthesized based on the coat protein coding region of tobacco ringspot vims (TRSV)for reverse transcription loop-mediated isothermal amplification ( RT-LAMP) assay. The amplification was detected vising the real time turbidimeter. An accelerated RT-LAMP procedure with one step was developed for the detection of TRSV. The specificity and sensitivity of the RT-LAMP assay were tested. RT-PCR compared to RT-LAMP by serial dilutions of total RNA extracts,they had the same sensitivities,but RT-LAMP was quicker,and the reaction could be finished within 1 hour. In addition,the presence or absence of the fluorescent display in daylight allowed naked easy detection of the amplification of TRSV genomic RNA using Calcein. The RT-LAMP assay was applied successfully to detect TRSV in soybean seeds,and the result by the addition of Calcein was consistent with the result detected by the real time turbidimeter. The method is rapid, specific, sensitive and suitable for rapid field detection of TRSV.关键词
烟草环斑病毒/逆转录环介导等温扩增/检测Key words
Tobacco ringspot virus (TRSV)/(Reverse transcription loop-mediated isothermal amplification ( RT-LAMP)/detection分类
农业科技引用本文复制引用
郭立新,陈先锋,徐亚飞,邱志君,朱迪琦,段维军..应用逆转录环介导等温扩增技术检测大豆种子中烟草环斑病毒[J].大豆科学,2012,31(6):980-983,987,5.基金项目
宁波检验检疫局科研项目(甬K07-2011) （甬K07-2011）
国家质检总局课题(2011IK169) （2011IK169）
质检公益性行业科研专项项目(201010256) （201010256）
浙江省重点科技创新团队项目(2010R50028). （2010R50028）
