应用复合RT-PCR同时检测烟草环斑病毒和番茄环斑病毒  

Establishment of Multiplex RT-PCR to Rapid Detection of Tobacco Ringspot Virus and Tomato Ringspot Virus

作　　者：吴兴海[1] 陈长法[1] 张云霞[1] 邵秀铃[1] 姜玉兰 

机构地区：[1]山东出入境检验检疫局,山东青岛266002 [2]青岛市植物保护站,山东青岛266002

出　　处：《江苏农业学报》2006年第4期398-400,共3页Jiangsu Journal of Agricultural Sciences

基　　金：国家质检总局植物病原检测专项基金(2005IK066)

摘　　要：将烟草环斑病毒(Tobacco ringspot virus)和番茄环斑病毒(Tom ato ringspot virus)的CP基因及RNA2基因序列引物,合成特异性RT-PCR检测引物,对烟草环斑病毒和番茄环斑病毒的感病植物组织进行了PCR扩增反应,结果显示,感染了烟草环斑病毒和番茄环斑病毒的植物组织的PCR产物均出现600 bp和470 bp特异性扩增条带,而其他病毒均未出现扩增条带,证明这两对引物具有烟草环斑病毒和番茄环斑病毒的鉴定特异性。将提取的烟草环斑病毒和番茄环斑病毒总RNA做梯度稀释,测定该检测体系的敏感度,结果显示,此体系最低可检出烟草环斑病毒和番茄环斑病毒提取的RNA模板浓度为234 pg/μl和264 pg/μl。表明,复合RT-PCR是一种可同时检测烟草环斑病毒和番茄环斑病毒的快速检测方法。Specific primers which were designed based on the sequences of Cp gene and RNA2 by multiplex RT-PCR were used to detect 5 virus, only in RNA extract from plant tissue infected with TRSV and TomRSV, the specific product of 600 bp and 470 bp DNA can be amplificd with their specific primers respectively . The lowest positive signal could be amplified from up to 234 pg/μl TRSV infected leaf total RNA and 264 pg/μl TomRSV infected leaf total RNA. RT-PCR method was showed to be spesific, sensitive , rapid for detction of TRSV and TomRSV.

关 键 词：复合RT-PCR 烟草环斑病毒 番茄环斑病毒 快速检测 

分 类 号：S41-30[农业科学—植物保护]