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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
【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。
【Abstract】 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.
【Key words】 multiplex RT-PCR; tobacco ringspot virus; tomato ringspot virus; rapid detection;
- 【文献出处】 江苏农业学报 ,Jiangsu Journal of Agricultural Sciences , 编辑部邮箱 ,2006年04期
- 【分类号】S432.41
- 【被引频次】7
- 【下载频次】225