Texas AgriLife Research and Extension Center, Amarillo, TX 79106, USA.
J Virol Methods. 2010 May;165(2):198-201. doi: 10.1016/j.jviromet.2010.01.019. Epub 2010 Feb 4.
Wheat streak mosaic virus (WSMV) and Triticum mosaic virus (TriMV) are widespread throughout the southwestern Great Plains states. When using conventional diagnostics such as enzyme-linked immunosorbent assays (ELISA), these two viruses are commonly found together in infected wheat samples. Methods for molecular detection have been developed for wheat viral pathogens, but until recently no multiplex method for detection of both WSMV and TriMV within a single sample was available. Therefore, the objective of this study was to develop a multiplex real-time PCR technique for detection of both pathogens within a single plant sample. Specific primers and probe combinations were developed for detection of WSMV and TriMV, single and multiple reactions were run simultaneously to detect any loss in sensitivity during the multiplex reaction, as well as any cross-reaction with other common wheat viruses. The multiplex reaction was successful in detection of both pathogens, with little difference between single and multiplex reactions, and no cross-reaction was found with other common wheat viruses. This multiplex technique not only will be useful for diagnostic evaluations, but also as a valuable tool for ecological and epidemiology studies, and investigations of host/pathogen interactions, especially when the host is infected with both pathogens.
小麦线条花叶病毒（WSMV）和小麦花叶病毒（TriMV）广泛存在于美国西南部大平原各州。当使用酶联免疫吸附测定（ELISA）等常规诊断方法时，这两种病毒通常在受感染的小麦样本中同时出现。已经开发出用于检测小麦病毒病原体的分子检测方法，但直到最近，还没有在单个样本中同时检测 WSMV 和 TriMV 的多重方法。因此，本研究的目的是开发一种用于在单个植物样本中同时检测这两种病原体的多重实时 PCR 技术。针对 WSMV 和 TriMV 开发了特异性引物和探针组合，同时进行单重和多重反应，以检测多重反应过程中是否存在灵敏度损失，以及是否与其他常见小麦病毒发生交叉反应。该多重反应成功地检测到了这两种病原体，单重和多重反应之间几乎没有差异，也没有与其他常见小麦病毒发生交叉反应。这种多重技术不仅将有助于诊断评估，而且还将成为生态和流行病学研究以及宿主/病原体相互作用研究的宝贵工具，特别是当宿主同时感染这两种病原体时。
