检测小麦线条花叶病毒的TaqMan MGB探针技术  

Detection of Wheat Streak Mosaic Virus Using TaqMan MGB Probe

作　　者：闻伟刚[1] 谭钟[1] 张吉红[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：《麦类作物学报》2009年第2期351-355,共5页Journal of Triticeae Crops

基　　金：宁波出入境检验检疫局科技项目(甬K01K-2008)

摘　　要：为给小麦线条花叶病毒(Wheat streak mosaic virus,WSMV)的灵敏、稳定、快速检测提供依据,根据该病毒不同分离株外壳蛋白(Coat protein,CP)基因的保守序列,设计了特异性引物与Taq Man MGB荧光探针,建立了WSMV的实时荧光RT-PCR检测方法。结果表明,本研究设计的Taq Man MGB荧光探针对WSMV的检测具有很好的特异性,解决了因病毒不同分离物之间基因组变异较大、难以找到长片段保守序列来设计探针的困难。该方法无需任何PCR后处理,基因污染风险小。它与双抗体夹心酶联免疫检测方法相比,灵敏度提高1 000倍,具有快速、灵敏和高特异性的优点,适合于WSMV的快速检测。For detection of wheat streak mosaic virus （WSMV）, a newly increased quarantine pest, a realtime fluorescent RT-PCR method was established with specific primers and TaqMan MGB probe, which were designed from the conservative sequences in coat protein genes of several WSMV isolates. Results showed that the TaqMan MGB probe was high specific for detection of WSMV, which solved the problem of probe designation without long conservative sequences among high genomic variability isolates. This technology obviates any post-PCR manipulation and reduces contamination risks. Compared to DASELISA method, the real-time fluorescent RT-PCR assay showed 1000-fold higher in detection sensitivity. It is a rapid, sensitive and highly specific method for detection of WSMV.

关 键 词：小麦线条花叶病毒 TAQMAN MGB探针 实时荧光RT—PCR 

分 类 号：S512.1[农业科学—作物学] S435.121.5