黄瓜花叶病毒荧光定量PCR检测方法的建立  

Real-time PCR for quantification of Cucumber mosaic virus in banana

作　　者：孙洁[1] 王婉[1] 周翎[1] 阮小蕾[1] 饶雪琴[1] 李华平[1] 

机构地区：[1]华南农业大学资源环境学院,广东广州510642

出　　处：《华南农业大学学报》2014年第2期53-56,共4页Journal of South China Agricultural University

基　　金：公益性行业(农业)科研专项(201203076-07)

摘　　要：【目的】建立检测香蕉中黄瓜花叶病毒Cucumber mosaic virus(CMV)的实时荧光定量方法.【方法】根据CMV外壳蛋白(CP)保守序列设计了TaqMan实时荧光定量PCR特异性探针及引物,优化反应体系检测TaqMan探针实时荧光定量方法的灵敏度、特异性和重复性.【结果和结论】该方法检测灵敏度为4.2×102μL-1,比普通PCR高100倍,且与香蕉束顶病毒Banana bunchy top virus(BBTV)、香蕉线条病毒Banana streak virus(BSV)无交叉反应,特异性和重复性都较好.用实时荧光定量PCR检测14份田间香蕉样品有5份样品为阳性,进一步证明建立的实时荧光定量方法可用于香蕉CMV的检测.[Objective] To develop a real-time quantitative PCR method with TaqMan probes to quantify Cucumber mosaic virus （CMV） in banana.[Method] The primers and probes were designed based on the conserved coat protein （ CP） sequences of CMV and were applied to real-time PCR assays .The reaction system was optimized ,and its sensitivity , specificity and repeatability were evaluated .[Result and conclu-sion]The detection sensitivity of the real-time PCR assay was 4.2 ×102μL-1 , which was 100 times more sensitive than PCR.The specifity of the assay was analyzed with Banana bunchy top virus （BBTV） and Banana streak virus （ BSV） , and no cross reaction were observed .The assay also had good repetitions . The real-time PCR assay was evaluated with field samples .5 of the 14 tissue samples collected from field suspected CMV infected bananas were positive , which further confirmed that the real-time PCR method should be suitable for detection and quantitation of CMV in banana .

关 键 词：黄瓜花叶病毒 TAQMAN探针 荧光定量PCR检测方法 

分 类 号：S436.421[农业科学—农业昆虫与害虫防治]