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

【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 conclusion】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.