[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 .%[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.
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