Institute of Plant Protection, Jiangsu Academy of Agricultural Sciences/Key Lab of Food Quality and Safety of Jiangsu Province - State Key Laboratory Breeding Base, Nanjing 210014, China.
Institute of Plant Protection, Jiangsu Academy of Agricultural Sciences/Key Lab of Food Quality and Safety of Jiangsu Province - State Key Laboratory Breeding Base, Nanjing 210014, China.
J Virol Methods. 2021 Dec;298:114277. doi: 10.1016/j.jviromet.2021.114277. Epub 2021 Sep 4.
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 × 10 µg 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.
烟草轻型绿斑病毒（TMGMV）是烟草花叶病毒属的一个成员种，感染辣椒（Capsicum annuum）和茄科的许多其他重要经济物种。TMGMV 感染严重影响了全球的辣椒生产，包括中国。本研究开发了一种用于检测辣椒田间样本和种子中 TMGMV 的反转录环介导等温扩增（RT-LAMP）检测方法。该方法基于与 TMGMV 基因组 C 末端区域的六个序列匹配的四个引物。RT-LAMP 检测法可从辣椒叶片的总 RNA 提取物中检测到病毒的存在，其灵敏度比相应的反转录聚合酶链反应（RT-PCR）检测法高 10 倍。该方法特异性检测 TMGMV，但不检测同属的密切相关种，如辣椒轻斑驳病毒、黄瓜绿斑驳花叶病毒和番茄花叶病毒。此外，SYBR Green I 的使用促进了 TMGMV RT-LAMP 产物的肉眼检测。这些结果表明，RT-LAMP 检测法是一种简单、敏感、特异和经济实惠的诊断工具，具有检测和监测田间样本中 TMGMV 感染的潜力。
