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根据GenBank中已发表的香蕉束顶病毒(Banana bunchy top virus,BBTV)﹑黄瓜花叶病毒(Cucumber mosaic virus,CMV)和香蕉线条病毒(Banana streak virus,BSV)的基因序列,找出保守序列分别设计出特异引物,在建立各种病毒单项PCR技术的基础上,通过优化多重PCR反应条件,建立了能同时检测3种病毒的多重PCR检测体系。该体系可同时扩增BBTV的615bp、CMV的480bp和BSV的945bp的特异片段。这些片段克隆测序结果表明,多重PCR扩增的不同病毒片段是特异和专化的,其核苷酸序列与相应的病毒基因片段的相似性都达91%以上。该体系的灵敏度测定结果表明,从相当于或大于10-2mg感病植物组织中均能检测到这3种病毒。
Based on the published gene sequences of Banana bunchy top virus (BBTV), Cucumber mosaic virus (CMV) and Banana streak virus (BSV) in GenBank, Based on the establishment of various single PCR techniques, a multiplex PCR system was established to detect three kinds of viruses by optimizing the conditions of multiplex PCR. This system can simultaneously amplify 615bp BBTV, 480bp CMV and 945bp specific fragment of BSV. The results of cloning and sequencing of these fragments showed that different virus fragments amplified by multiplex PCR were specific and specific, and their nucleotide sequences were more than 91% similar to the corresponding viral gene fragments. The sensitivity of the system showed that all three viruses could be detected from the tissues corresponding to or more than 10-2 mg of susceptible plants.