Development of Multiplex RT-PCR for Simultaneous Detection of Three Viruses in Bulbous Iris
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摘要: 根据GenBank中已发表的鸢尾轻花叶病毒(Iris mild mosaic virus,IMMV)、水仙潜隐病毒(Narcissus latent virus,NLV)和菜豆黄花叶病毒(Bean yellow mosaic virus,BYMV)外壳蛋白(CP)基因序列保守区域分别设计特异性引物,通过优化多重PCR反应条件,建立能同时检测球根鸢尾3种病毒的多重PCR检测体系。该体系能够一次扩增出IMMV、NLV和BYMV的特异片段,其大小分别是770bp、266bp和186bp。测序结果表明,3种病毒序列与相应的参考序列相似性均达98%以上。灵敏度测定结果表明,从相当于或大于10-2 mg的感病植物组织中能够检测到这3种病毒。Abstract: By using specific primers designed on the basis of conserved sequences of coat protein(CP)gene,a multiplex PCR protocol for the detection of three main viruses(Iris mild mosaic virus,IMMV;Narcissus latent virus,NLV;Bean yellow mosaic virus,BYMV)in Bulbous Iris was developed.Three specific fragments were simultaneously amplified in one PCR reaction system.Their molecular weights were 770 bp,266bp and186 bp respectively.Sequence analysis indicated that the three virus sequences shared at least 98 percent identities with that of other relative viruses.The result showed that the sensitivity of this protocol equaled to be able to detect the three viruses at the level of 10-2 mg Bulbous Iris tissue.
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