CAJ | 학술논문

[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．
[목적]건립검측향초중황과화협병독Cucumber mosaic virus（ CMV）적실시형광정량방법．[방법]근거CMV외각단백（ CP）보수서렬설계료TaqMan실시형광정량PCR특이성탐침급인물，우화반응체계검측TaqMan탐침실시형광정량방법적령민도、특이성화중복성．[결과화결론]해방법검측령민도위4.2×102μL-1，비보통PCR고100배，차여향초속정병독Banana bunchy top virus（BBTV）、향초선조병독Banana streak virus（BSV）무교차반응，특이성화중복성도교호．용실시형광정량PCR검측14빈전간향초양품유5빈양품위양성，진일보증명건립적실시형광정량방법가용우향초CMV적검측．
[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 .