One-step reverse transcription loop-mediated isothermal amplification for the rapid detection of cucumber green mottle mosaic virus

文献类型： 外文期刊

第一作者： Wei, Qi-wei

作者： Wei, Qi-wei;Zhang, Wen-na;Wu, Jian-yan;Charimbu, Miriam Karwitha;Hu, Bai-shi;Tao, Xiao-rong;Liu, Yong;Tan, Xin-qiu;Hu, Bai-shi;Cheng, Zhao-bang;Yu, Cui

作者机构：

关键词： Cucumber green mottle mosaic virus;RT-LAMP;Watermelon;Seed

期刊名称：JOURNAL OF VIROLOGICAL METHODS （ 影响因子：2.014； 五年影响因子：2.001 ）

ISSN： 0166-0934

年卷期： 2013 年 193 卷 2 期

页码：

收录情况： SCI

摘要： Cucumber green mottle mosaic virus (CGMMV) has caused serious damage, to Cucurbitaceae crops worldwide. The virus is considered one of the most serious Cucurbitaceae quarantine causes in many countries. In this study, a highly efficient and practical one-step reverse transcription loop-mediated isothermal amplification (RT-LAMP) was developed for the detection of CGMMV. The total RNA or crude RNA extracted from watermelon plants or seeds could be detected easily by this RT-LAMP assay. The RT-LAMP assay was conducted in isothermal (63 degrees C) conditions within 1 h. The amplified products of CGMMV could be detected as ladder-like bands using agarose gel electrophoresis or visualized in-tube under UV light with the addition of a fluorescent dye. The RT-LAMP amplification was specific to CGMMV, as no cross-reaction was observed with other viruses. The RT-LAMP assay was 100-fold more sensitive than that of reverse-transcription polymerase chain reaction (RT-PCR). This is the first report of the application of the RT-LAMP assay to detect CGMMV. The sensitive, specific and rapid RT-LAMP assay developed in this study can be applied widely in laboratories, the field and quarantine surveillance of CGMMV. (C) 2013 Elsevier B.V. All rights reserved.

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