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香蕉苞片花叶病毒(Banana bract mosaic virus,BBrMV)是我国对外检疫性有害生物。为防止该有害生物传入我国,根据BBrMV不同分离株外壳蛋白基因(coat protein,CP)的保守序列,设计特异性引物与TaqMan荧光探针,建立了BBrMV的实时荧光RT-PCR检测方法。结果表明,该方法检测BBrMV的2个不同来源毒株,均能够得到典型扩增曲线,Ct值分别为26.65和27.52;而马铃薯Y病毒、李属坏死环斑病毒以及桃丛簇花叶病毒等其它毒株则没有典型扩增曲线,也无Ct值。灵敏度比较发现,该方法比普通RT-PCR检测方法的灵敏度提高1000倍,具有快速、灵敏和高特异性的优点,适合对BBrMV的检测。
Banana bract mosaic virus (BBrMV) is an external quarantine pest in China. To prevent the introduction of this pest into our country, a real-time fluorescent RT-PCR method for the detection of BBrMV was established based on the conserved sequence of the coat protein (CP) gene of different BBrMV isolates. The specific primers and TaqMan probe were designed. The results showed that two typical strains of BBrMV could detect typical amplification curves with Ct values of 26.65 and 27.52, respectively. However, potato Y virus, Prunus necrosis ring spot virus and peach mosaic virus Other strains do not have typical amplification curves nor Ct values. Sensitivity comparison found that this method than ordinary RT-PCR detection method to improve the sensitivity of 1000 times, with the advantages of rapid, sensitive and high specificity, suitable for the detection of BBrMV.