Development of a reverse-transcription loop-mediated isothermal amplification assay for the detection of Tobacco mild green mosaic virus (TMGMV)

文献类型： 外文期刊

第一作者： Zhao, Wenhao

作者： Zhao, Wenhao;Wu, Shuhua;Du, Linlin;Li, Tingfang;Cheng, Zhaobang;Zhou, Yijun;Ji, Yinghua

作者机构：

关键词： Tobacco mild green mosaic virus (TMGMV); Reverse-transcription loop-mediated; isothermal amplification (RT-LAMP); Virus detection; Pepper; Capsicum annuum

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： 2021 年 298 卷

页码：

收录情况： SCI

摘要： Tobacco mild green mosaic virus (TMGMV), a member species of the genus Tobamovirus, infects pepper (Capsicum annuum) and a number of other economically important species in the Solanaceae family. TMGMV infections had seriously impacted pepper production worldwide, including China. A reverse-transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed to detect TMGMV in pepper field samples and seed. This assay was based on four primers that matched to six sequences in the C-terminal region of the TMGMV genome. RT-LAMP assay could detect the presence of the virus in 3.0 x 10- 7 mu g of total RNA extract from pepper leaves, which was ten times more sensitive than the corresponding reverse-transcription polymerase chain reaction (RT-PCR) assay. This method specifically detected TMGMV but not the closely related species of the same genus Pepper mild mottle virus, Cucumber green mottle mosaic virus and Tomato mosaic virus. In addition, the use of SYBR Green I facilitated the detection of the TMGMV RT-LAMP products by the naked eye. These results indicated that the RT-LAMP assay was a simple, sensitive, specific and affordable diagnostic tool that has the potential to detect and monitor TMGMV infection in field samples.

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