基因芯片技术检测黄瓜花叶病毒、烟草花叶病毒和马铃薯Y病毒

摘要

The Cucumber mosaic virus ( CMV), Tobacco mosaic virus (TMV) and Potato virus Y ( PVY ) are significant worldwide plant viruses,which harm to agricultural production. Mixed infection with several viruses usually occurs complex symptom. So, rapid,sensitive and specific method,by which several viruses can be detected simultaneously,is needed urgently. In this study, according to three kinds of viral coat protein gene sequence,primers and probes were designed,gene chip was prepared. The downstream primers were labled by Cy3. The product amplified by RT-PCR using the labled primers was hybridized with the gene chip. The hybridization signal was detected and analyzed by a fluorescence scanner. The result showed that the specific signals were identified from virus-infected plants.The detection sensitivities of gene chip were 10-100 times higher than RT-PCR. The results indicate that gene chip can be used for detection of plant viruses fast and accurately.%黄瓜花叶病毒、烟草花叶病毒和马铃薯Y病毒是世界性分布的重要植物病毒,对农业生产危害较大.农作物感病多为复合侵染,症状表现较为复杂,急需建立快速、准确、能够一次检测多种病毒的检测方法.本研究根据3种病毒的外壳蛋白基因序列,设计引物和探针,制备基因芯片;用Cy3标记下游引物,RT-PCR扩增产物与芯片杂交,荧光扫描仪检测并分析信号.结果表明,该芯片可以从病毒侵染样本中检测到特异性识别信号,检测灵敏度比RT-PCR高10~100倍,该技术能对植物病毒做出快速、准确的检测.