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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。
Grapevine leafroll virus strain Ⅲ was detected by RT-PCR. The technological process adopted is: obtaining RNA of the virus by extracting total RNA of grape leaves or phloem, reverse transcribing and synthesizing cDNA, obtaining characteristic fragments of virus cDNA by PCR amplification and carrying out sequence analysis. The integrity and purity of the extracted total RNA were tested, and a good RNA acquisition method was determined. The results showed that the homology of the fragment amplified by RT-PCR with the virus source sequence was as high as 99.3%, indicating that the determination of grape leafroll virus strain III using the method identified in this study is reliable.