基于TaqMan MGB探针的李痘病毒分子检测  

Molecular detection of Plum pox virus based on TaqMan MGB probe

作　　者：闻伟刚[1] 谭钟[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：《果树学报》2009年第4期581-584,共4页Journal of Fruit Science

基　　金：宁波出入境检验检疫局科技项目(甬K01-2009)

摘　　要：李痘病毒(Plum pox virus,PPV)是我国对外检疫性有害生物。研究根据该病毒不同分离株外壳蛋白(coatprotein,CP)基因的保守序列,设计了特异性引物与荧光探针,建立了基于TaqMan MGB分子探针的PPV实时荧光RT-PCR检测方法。方法特异性研究表明,针对PPV的2个主要流行株系M株系与D株系,均能够得到典型扩增曲线,Ct值分别为14.96和18.22;而对于马铃薯Y病毒、李属坏死环斑病毒以及桃丛簇花叶病毒等其他毒株则没有典型扩增曲线,也无Ct值。灵敏度比较发现,该方法比双抗体夹心酶联免疫检测方法的灵敏度提高1000倍,具有快速、灵敏和高特异性的优点,适合对PPV的检测。Plum pox virus （PPV） is a quarantine disease in China. In this study, based on conservative sequences of the coat protein genes of different PPV isolates, a real-time fluorescent RT-PCR method was established with specific primers and TaqMan MGB probe. Specific studies have shown that for the two main epidemic strains M and D of PPV, a typical amplification curve can be obtained with Ct values of 14.96 and 18.22; and for the potato Y virus ,Prunus necrotic ringspot virus, peach rosette mosaic virus and other virus strains, typical amplification curve and Ct values could not be obtained. Sensitivity of this method was 1000 times higher than that of double-antibody sandwich enzyme-linked immunosorbent assay. Thus, the real-time fluorescent RT-PCR was a rapid, sensitive and highly specific method for the detection of PPV.

关 键 词：李痘病毒 TAQMAN MGB探针 实时荧光RT-PCR 检测 

分 类 号：S662.3[农业科学—果树学]