甘薯羽状斑驳病毒和褪绿矮化病毒双重RT-PCR检测方法的建立

Development of Dutiplex RT-PCR for the Detection of SPFMV and SPCSV in Sweet Potato

摘要: 根据GenBank中公布的甘薯羽状斑驳病毒 (Sweet potato feathery mottle virus, SPFMV) 外壳蛋白 (CP) 基因和甘薯褪绿矮化病毒 (Sweet potato chlorotic stunt virus, SPCSV) 热激蛋白 (Hsp70) 基因序列保守区域设计了2对特异性引物, 通过RT-PCR反应程序的优化, 建立了能同时检测SPFMV和SPCSV的双重PCR检测体系。该检测体系能够1次扩增出SPFMV和SPCSV的特异性片段, 片段大小为461bp和304bp, 测序结果表明, 2种病毒序列与参考序列的同源性达93%以上。

Abstract: A duplex PCR protocol for the detection of two main viruses in sweet potato was developed based on the coat protein (CP) gene of Sweet potato feathery mottle virus (SPFMV) and the (Hsp70) gene of Sweet potato chlorotic stunt virus (SPCSV) .They were able to amplify unique DNA fragments of 461 bp and 304 bp in a single PCR reaction.The sequence analysis suggst that the two viruses shared at least 93 percent homology with other related viruses.