中文摘要：小麦矮缩病毒（WDV）、大麦黄条点花叶病毒（BYSMV）、水稻黑条矮缩病毒（RBSDV）和北方禾谷花叶病毒（NCMV）是四种侵染小麦且致病症状相似的病毒。本文采用多重逆转录聚合酶链反应（m-RT-PCR）法对这四种病毒进行同步检测与识别。针对每个病毒设计了不同的特异性引物，并生成了四个不同的片段（273、565、783和1296 bp），分别检测RBSDV、BYSMV、WDV和NCMV的存在。对m-RT-PCR退火温度、dNTP浓度、Taq聚合酶和Mg2+进行优化。通过来自中国不同地区（已发现RBSDV、BYSMV、WDV）的田间样本对引物扩增特异性进行检测。本研究实现了小麦病毒的快速、特异性检测，并为调查这些新病毒病的流行病学提供了可能性。
外文摘要：Wheat dwarf virus (WDV), barley yellow striate mosaic virus (BYSMV), rice black-streaked dwarf virus (RBSDV) and northern cereal mosaic virus (NCMV) are four viruses infecting wheat and causing similar symptoms. In this paper, a multiplex reverse transcription polymerase chain reaction (m-RT-PCR) method has been developed for the simultaneous detection and discrimination of these viruses. The protocol uses specific primer set for each virus and produces four distinct fragments (273, 565, 783 and 1296 bp), detecting the presence of RBSDV, BYSMV, WDV and NCMV, respectively. Annealing temperature, concentrations of dNTP, Taq polymerase and Mg2+ were optimized for the m-RT-PCR. The detection limit of the assay was up to 10(-2) dilution. The amplification specificity of these primers was tested against a range of field samples from different regions of China, where RBSDV, BYSMV, WDV have been detected. This study fulfills the need for a rapid and specific wheat virus detection that also has the potential for investigating the epidemiology of these new viral diseases.
外文关键词：Wheat viruses；RNA virus；DNA virus；Multiplex RT-PCR
作者：Zhang, Peipei；Liu, Yan；Liu, Wenwen；等
作者单位：中国农业科学院
期刊名称：JOURNAL OF VIROLOGICAL METHODS
期刊影响因子：1.6929
出版年份：2017
出版刊次：11
点击下载：多重RT-PCR法同步检测小麦矮缩病毒、北方禾谷花叶病毒、大麦黄条点花叶病毒和水稻黑条矮缩病毒的研究