实时荧光RT-PCR方法检测黄瓜绿斑驳花叶病毒  

Detection of Cucumber green mottle mosaic virus by real time Reverse Transcriptase Polymerase Chain Reaction

作　　者：邓丛良[1] 黄峰[2] 吕玉峰[1] 张凯[1] 郭育林 赵焕生 

机构地区：[1]北京出入境检验检疫局植物检疫中心,北京100026 [2]厦门出入境检验检疫局 [3]北京金纳信生物科技有限公司

出　　处：《植物检疫》2009年第4期29-31,共3页Plant Quarantine

基　　金："十一五"支撑项目(2006BAK10B06);国家质检总局课题(2008IK248)资助

摘　　要：根据黄瓜绿斑驳花叶病毒不同分离物外壳蛋白基因(CP)的保守序列,设计合成引物和TaqMan荧光探针,建立了黄瓜绿斑驳花叶病毒的实时荧光RT-PCR(Real time RT-PCR)检测方法。该方法利用TaqMan探针水解产生的荧光信号实时监测目标基因的扩增,实现PCR扩增与检测同步进行。结果表明,实时荧光RT-PCR方法检测灵敏度比普通RT-PCR高约10倍,并且具有快速、简便、准确的优点,适合于CGMMV的快速、高效检测。A pair of primers and a TaqMan probe were designed based on the conserved nucleotide sequence of coat protein （CP） gene of known Cucumber green mottle mosaic virus （CGMMV） isolates. The real time fluorescent RT - PCR system was developed for the detection of CGMMV, in which accumulation of fluorescence signal and amplification of the target gene could be monitored simultaneously. The method presented here was 10 times more sensitive than conventional RT - PCR , allowing rapid and reliable detection of CGMMV.

关 键 词：黄瓜绿斑驳花叶病毒 实时荧光RT—PCR 检测 

分 类 号：S41[农业科学—植物保护]