State Key Laboratory of Crop Stress Biology in Arid Areas and Key Laboratory of Crop Pest Integrated Pest Management on the Loess Plateau of Ministry of Agriculture, College of Plant Protection, Northwest A&F University, Yangling, 712100, China.
J Virol Methods. 2012 Jul;183(1):57-62. doi: 10.1016/j.jviromet.2012.03.029. Epub 2012 Mar 31.
Tobacco viruses including Tobacco mosaic virus (TMV), Cucumber mosaic virus (CMV), Tobacco etch virus (TEV), Potato virus Y (PVY) and Tobacco vein banding mosaic virus (TVBMV) are major viruses infecting tobacco and can cause serious crop losses. A multiplex reverse transcription polymerase chain reaction assay was developed to detect simultaneously and differentiate all five viruses. The system used specific primer sets for each virus producing five distinct fragments 237, 273, 347, 456 and 547 bp, representing TMV, CMV subgroup I, TEV, PVY(O) and TVBMV, respectively. These primers were used for detection of the different viruses by single PCR and multiplex PCR and the results were confirmed by DNA sequencing analysis. The protocol was used to detect viruses from different parts of China. The simultaneous and sensitive detection of different viruses using the multiplex PCR is more efficient and economical than other conventional methods for tobacco virus detection. This multiplex PCR provides a rapid and reliable method for the detection and identification of major tobacco viruses, and will be useful for epidemiological studies.
烟草病毒包括烟草花叶病毒（TMV）、黄瓜花叶病毒（CMV）、烟草蚀纹病毒（TEV）、马铃薯 Y 病毒（PVY）和烟草脉带斑驳病毒（TVBMV），是侵染烟草的主要病毒，可以引起严重的作物损失。建立了一种同时检测和区分这五种病毒的多重反转录聚合酶链式反应（RT-PCR）检测方法。该系统使用针对每种病毒的特异性引物组，产生 237、273、347、456 和 547bp 的 5 个不同片段，分别代表 TMV、CMV 亚组 I、TEV、PVY（O）和 TVBMV。这些引物用于单重 PCR 和多重 PCR 检测不同病毒，并用 DNA 测序分析进行验证。该方案用于检测来自中国不同地区的病毒。与其他烟草病毒检测的常规方法相比，该多重 PCR 具有更高的效率和经济性，可同时灵敏地检测不同的病毒。该多重 RT-PCR 为主要烟草病毒的检测和鉴定提供了一种快速可靠的方法，将有助于进行流行病学研究。
