双重RT-PCR同步检测马铃薯A病毒和马铃薯卷叶病毒  

Research on the detection of PVA and PLRV by Duplex RT-PCR

作　　者：宁红 张涛[2] 孟兴[3] 张敏[2] 郭迪金 万佳 

机构地区：[1]四川省植物检疫站,成都610041 [2]四川农业大学雅安 [3]四川出入境检验检疫局成都

出　　处：《植物检疫》2009年第5期28-30,共3页Plant Quarantine

基　　金：四川省科技攻关项目(07FG002-008)

摘　　要：根据马铃薯A病毒全基因序列和马铃薯卷叶病毒衣壳蛋白区序列分别设计PVA和PLRV的特异性引物,应用双重RT-PCR同步检测马铃薯A病毒和马铃薯卷叶病毒,分别得到300bp和222bp大小的扩增片段。试验从反转录、PCR等方面对双重RT-PCR同步检测2种病毒进行了探索和优化。结果表明反转录反应中dNTPs浓度、2种病毒下游引物浓度比例以及PCR反应中Mg2+浓度对双重RT-PCR同时检测PVA和PLRV有较大的影响。A duplex reverse transcription polymerase chain reaction was developed for the simultaneous detection of PVA and PLRV. The primers were designed based on the complete genome of PVA and the coat protein gene of PLRV, and their amplification fragments by RT -PCR were 300bp （PVA） and 222bp （PLRV）. Reverse transcription （RT） stage and polymerase chain reaction （PCR） stage were researched and optimized in this experiment. The result showed that the dNTPs concentration, antisense primer rations of two viruses at RT stage and Mg^2＋ concentration on PCR stage had a great impact on the detection.

关 键 词：双重RT-PCR 马铃薯 病毒检测 

分 类 号：S435.32[农业科学—农业昆虫与害虫防治]