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

摘要

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 304bp in a single PCR reaction.The sequence analysis suggst that the two viruses shared at least 93 percent homology with other related viruses.%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461 bp 和304 bp，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。