根据香蕉束顶病毒(Banana bunchy top virus,BBTV)复制酶基因保守序列设计特异性探针及引物,建立了BBTV实时荧光PCR(Taq Man real-time PCR)检测方法。结果显示,所建立的检测方法在检测质粒DNA标准品和发病香蕉植株总DNA时,其灵敏度均比普通PCR高,且与黄瓜花叶病毒(Cucumber mosaic virus,CMV)和香蕉线条病毒(Banana streak virus,BSV)无交叉反应。同浓度样品进行7次重复性试验,各重复扩增结果所得Ct值的变异系数为0.93%,表明建立的荧光定量PCR检测方法重复性好,Ct值保持稳定。利用所建立的方法检测带毒香蕉吸芽繁殖的组培苗中的BBTV,发现BBTV含量随着继代数的增加而增加,并且组培苗顶部叶片中的含量高于其他部位的叶片。 According to the conserved sequence of replicase gene of Banana bunchy top virus (BBTV), specific probes and primers were designed and used to detect BBTV real-time PCR (Taq Man real-time PCR). The results showed that the sensitivity of the established method was higher than that of ordinary PCR in the detection of plasmid DNA standard and the total DNA of diseased banana plants, and the sensitivity of the method to that of Cucumber mosaic virus (CMV) and banana streak virus, BSV) No cross reaction. The results of seven replicate experiments with the same concentration of samples showed that the coefficient of variation (Ct) was 0.93%. The results showed that the established method of fluorescence quantitative PCR had good repeatability and stable Ct value. The BBTV content in the tissue culture seedlings infected with banana sprouts was detected by the established method. It was found that the content of BBTV increased with the increase of the succession number, and the content of BBTV in the top leaves of the tissue culture seedlings was higher than that of other parts.