建立一种用于检测玉米中的玉米黄斑驳病毒的 RT-LAMP 检测方法。
Development of an RT-LAMP assay for the detection of maize yellow mosaic virus in maize.
机构信息
School of Life Science, Biocontrol Engineering Research Center of Crop Diseases & Pests, Yunnan Province, Biocontrol Engineering Research Center of Plant Diseases & Pests, Yunnan University, Kunming, 650091, China.
Yunnan Plant Protection and Quarantine Station, Kunming, 650034, China.
出版信息
J Virol Methods. 2022 Feb;300:114384. doi: 10.1016/j.jviromet.2021.114384. Epub 2021 Nov 29.
Maize is one of the most widely cultivated cereal crops worldwide. Maize yellow mosaic virus (MaYMV) (species Maize yellow mosaic virus, genus Polerovirus and family Luteoviridae) was first reported in maize from China. In this study, a one-step reverse-transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for detecting MaYMV. The optimal concentrations of betaine, Mg and dNTPs for the assay were 0 M, 1.4 mM and 6 mM, respectively, and the optimal reaction time was 50 min. Using total plant RNA as the template, the detection limit of the RT-LAMP assay for MaYMV was 1 pg, while that of RT-PCR was 100 pg, indicating that the RT-LAMP assay developed was 100 times more sensitive than RT-PCR. Importantly, the RT-LAMP assay successfully detected MaYMV using rapidly extracted crude RNA from infected maize as a template. In conclusion, the RT-LAMP assay developed was a rapid, specific, sensitive and low-cost method for the detection of MaYMV in field samples of maize.
玉米是世界上种植最广泛的谷物之一。玉米黄斑驳病毒（MaYMV）（种为玉米黄斑驳病毒，属为马铃薯 Y 病毒属，科为杆状病毒科）最初在中国的玉米中被报道。在本研究中，开发了一种一步逆转录环介导等温扩增（RT-LAMP）检测方法来检测 MaYMV。该测定法的最佳甜菜碱、Mg 和 dNTPs 浓度分别为 0 M、1.4 mM 和 6 mM，最佳反应时间为 50 min。使用总植物 RNA 作为模板，RT-LAMP 测定法检测 MaYMV 的检测限为 1 pg，而 RT-PCR 的检测限为 100 pg，表明 RT-LAMP 测定法比 RT-PCR 灵敏 100 倍。重要的是，该 RT-LAMP 测定法成功地使用从感染玉米中快速提取的粗 RNA 作为模板检测到 MaYMV。总之，该 RT-LAMP 测定法是一种快速、特异、灵敏且低成本的方法，可用于玉米田间样本中 MaYMV 的检测。
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