实时荧光RT-PCR检测香蕉苞片花叶病毒方法的建立  

Detection of Banana bract mosaic virus by real-time fluorescent RT-PCR

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,宁波315012

出　　处：《植物保护学报》2009年第5期417-420,共4页Journal of Plant Protection

基　　金：宁波出入境检验检疫局科技项目(甬K01-2009)

摘　　要：香蕉苞片花叶病毒(Banana bract mosaic virus,BBrMV)是我国对外检疫性有害生物。为防止该有害生物传入我国,根据BBrMV不同分离株外壳蛋白基因(coat protein,CP)的保守序列,设计特异性引物与TaqMan荧光探针,建立了BBrMV的实时荧光RT-PCR检测方法。结果表明,该方法检测BBrMV的2个不同来源毒株,均能够得到典型扩增曲线,Ct值分别为26.65和27.52;而马铃薯Y病毒、李属坏死环斑病毒以及桃丛簇花叶病毒等其它毒株则没有典型扩增曲线,也无Ct值。灵敏度比较发现,该方法比普通RT-PCR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BBrMV的检测。Banana bract mosaic virus （BBrMV） is a quarantine pests issued by Chinese government. To prevent the spread of the pests, a real-time fluorescent RT-PCR method was established based on specific primers and TaqMan probe which designed with the conservative sequences of the coat protein genes of different BBrMV isolates. Specificity study showed that typical amplification curves could be obtained for the two BBrMV strains, with Ct values of 26.65 and 27.52 ; and no typical amplification curve obtained for Potato Y virus, Plum pox virus, Prunus necrotic ringspot virus, Peach rosette mosaic virus and other viruses. The sensitivity of this method was 1000 times higher than of regular RT-PCR. Thus, the real-time fluorescent RT-PCR is a rapid, sensitive and highly specific method for the detection of BBrMV.

关 键 词：香蕉苞片花叶病毒 TAQMAN探针 实时荧光RT—PCR 检测 

分 类 号：S436.68[农业科学—农业昆虫与害虫防治]