摘 要 ：根据菜豆荚斑驳病毒(Bean pod mottle virus, BPMV)不同分离株外壳蛋白基因(coat protein, CP)的保守序列,设计了特异性引物与TaqMan MGB荧光探针,建立了BPMV的实时荧光RT-PCR检测方法。该方法与酶联免疫检测方法相比,灵敏度提高了100倍,具有快速、灵敏和高特异性的优点,适于对BPMV的快速检测。应用该方法,对来自美国不同地区的携带有BPMV的大豆样品进行检测,均能够得到BPMV的典型扩增曲线,表明引物与探针对BPMV不同分离株具有良好的简并性。
Abstract:A real-time fluorescent RT-PCR method for detection of Bean pod mottle virus (BPMV) was established with specific primers and TaqMan MGB probe, which were designed from the conservative sequences in coat protein genes of several BPMV isolates. Compared to ELISA method, the real-time fluorescent RT-PCR assay improved sensitivity by up to 100-fold. It is a rapid, sensitive and highly specific method for detection of BPMV. Using with this method, we obtained typical amplified curve from all the American soybean samples which infected by BPMV. The results showed that the primers and TaqMan MGB probe designed in this study were degenerate for various BPMV isolates.