关键词：
桑脉带相关病毒;反转录环介导等温扩增技术;检测方法
摘 要：
桑脉带相关病毒Mulberry vein banding-associated virus (MVBaV)是广西桑树病毒病的主要病原病毒。本研究根据MVBaV基因组的核外壳蛋白(nucleocapsid protein, N)基因序列设计了5组引物,筛选获得了1组有效引物,建立了该病毒的反转录环介导等温扩增技术(reverse transcription loop-mediated isothermal amplification, RT-LAMP)检测方法。该方法能在恒温条件(63℃)下1 h内检测MVBaV,其灵敏度是RT-PCR检测方法的10倍。对6个田间样品进行RT-LAMP检测,其结果与RT-PCR的结果一致。该方法可直接在反应管中加入SYBR GreenⅠ,通过颜色变化即可判定结果,无需经过琼脂糖电泳或专门仪器,具有灵敏、快速和特异性好的优点,可应用于MVBaV的快速诊断及其田间监测。
译 名：
Development of reverse transcription loop-mediated isothermal amplification for the detection of Mulberry vein banding-associated virus
作 者：
WU Fan;LI Yangxiu;HUANG Haijuan;CHEN Baoshan;MENG Jiaorong;College of Agriculture, Guangxi University;College of Life Science and Technology, Guangxi University;State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources;
单 位：
WU Fan%LI Yangxiu%HUANG Haijuan%CHEN Baoshan%MENG Jiaorong%College of Agriculture, Guangxi University%College of Life Science and Technology, Guangxi University%State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources
关键词：
mulberry vein banding-associated virus;;RT-LAMP;;detection method
摘 要：
Mulberry vein banding-associated virus(MVBaV) is one of the main pathogenic viruses infecting mulberry in Guangxi. Five sets of specific primers were designed based on MVBaV nucleocapsid protein(N) gene for reverse transcription loop-mediated isothermal amplification(RT-LAMP) assay, and one feasible set of primers was selected. The RT-LAMP method was developed for the detection of MVBaV from mulberry. In the RT-LAMP method, MVBaV genomic RNA could be amplified under isothermal conditions(63℃) within 1 h, and was 10 times more sensitive than RT-PCR. Consistent results of six mulberry samples from the field were obtained by the RT-LAMP and RT-PCR assay, suggesting the reliability of RT-LAMP in MVBaV detection.The results were validated by the measurement or observation of the color of the reaction mixture after adding SYBR GreenⅠwithout gel electrophoresis or special equipments. This RT-LAMP method enabled sensitive, rapid and specific detection of MVBaV and could be applied for the rapid diagnosis and surveillance of MVBaV in the field.
