Position: Home > Articles > Detection of Grapevine Leafroll-associated Virus Ⅲ by Reverse Tran-scription-polymerase Chain Reaction
Journal of Fruit Science
2002,19
(1)
15-18
葡萄卷叶病毒Ⅲ RT-PCR检测技术研究
作 者：
刘三军;郭卫东;张秋叶;何水涛;周厚成;张亚冰
单 位：
中国农业科学院郑州果树研究所;西北农林科技大学
关键词：
葡萄卷叶病毒Ⅲ;RT－PCR;检测;克隆;序列分析
摘 要：
利用ＲＴ－ＰＣＲ技术,对葡萄卷叶病毒株系Ⅲ进行了检测。采用的技术流程为:通过提取葡萄叶片或韧皮部总ＲＮＡ,获得病毒的ＲＮＡ,反转录合成ｃＤＮＡ,经ＰＣＲ扩增,获得病毒ｃＤＮＡ的特征片段,并进行了序列分析。对提取的总ＲＮＡ进行了完整性和纯度检测,确定了良好的ＲＮＡ获得方法。结果表明,通过ＲＴ－ＰＣＲ扩增的片段序列与病毒源序列的同源性高达９９．３%,说明采用本研究确定的方法检测葡萄卷叶病毒株系Ⅲ结果可靠。
译 名：
Detection of Grapevine Leafroll-associated Virus Ⅲ by Reverse Tran-scription-polymerase Chain Reaction
作 者：
Liu Sanjun,Zhang Yabing,Guo Weidong,Zhang Qiuye,He Shuitao,and Zhou Houcheng(Zhengzhou Fruit Research Institute,CAAS Zhengzhou,Henan 450009;Northwest Sci-Tech University of Agriculture and Forestry,Yangling,Shaanxi 712100)
关键词：
Grapevine leafroll-associated virus type Ⅲ;RT-PCR;Detection;Clone;Sequence analysis
摘 要：
An efficient procedure for the extraction of high-quality RNA from grape leaf and bark is described.A stable proce-dure for detection of grapevine leafroll-associated virus type Ⅲ was set up by the Reverse Transcription-Polymerase ChainReaction.The main technique is the method of the extraction of total RNA from grapevine leaves or bark.A 300 bp fragmentwas acquired by RT-PCR and the sequence of this fragment was aligned with the original sequence of grapevine leafroll-as-sociated virus type Ⅲ.The result showed that the identity of two sequence reach 99.3%.This indicated that the detection resultwas reliable.
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