玉米褪绿斑驳病毒实时荧光RT-PCR检测方法研究  

Detection of Maize chlorotic mottle virus by real-time fluorescent RT-PCR

作　　者：闻伟刚[1] 张建成[1] 崔俊霞[1] 张颖[1] 

机构地区：[1]宁波出入境检验检疫局,浙江宁波315012

出　　处：《植物检疫》2011年第1期39-42,共4页Plant Quarantine

基　　金：浙江省分析测试项目(2009F70036);质检公益性项目(201010256)

摘　　要：玉米褪绿斑驳病毒(Maize chlorotic mottle virus,MCMV)是我国对外公布的检疫性有害生物。本研究根据该病毒外壳蛋白基因的保守序列,设计得到特异性引物及Taqman荧光探针,建立了MCMV的实时荧光RT-PCR方法,并对其灵敏度与特异性进行了研究。该方法针对2个不同来源的毒株均能得到典型扩增曲线,而没有从小麦线条花叶病毒、玉米粗缩病毒和玉米矮花叶病毒的RNA得到扩增曲线,表明引物与荧光探针具有良好的特异性。针对玉米褪绿斑驳病毒RNA不同稀释度样品,实时荧光RT-PCR检测低限达到10-5稀释度,检测灵敏度要比普通RT-PCR高出100倍。因此,本研究建立的MCMV实时荧光方法具有特异性强、灵敏度高和快速有效的优点。Maize chlorotic mottle virus （MCMV） is a quarantine pests issued by Chinese government. In this study, accord to the conservative sequences of the coat protein genes of different MCMV isolates, a real - time flu- orescent RT- PCR method was established based on specific primers and TaqMan probe. Specific studies have shown that for the two MCMV strains from different sources, a typical amplification curve can be obtained, and not for the Wheat streak mosaic virus, Maize rough dwarf virus and Maize dwarf mosaic virus. Sensitivity compari- son found that the detection limit can reach 10-5 dilution, which is 100 times higher than the regular RT -PCR method. Thus, the real - time fluorescent RT - PCR is a rapid, sensitive and highly specific method for the detec- tion of MCMV.

关 键 词：玉米褪绿斑驳病毒 实时荧光RT—PCR 检测 

分 类 号：S432.41[农业科学—植物病理学]