摘 要 ：针对进口大豆的菜豆荚斑驳病毒(Bean pod mottle virus, BPMV)检测,建立了一步法RT-PCR和一步法实时荧光RT-PCR检测方法。依据BPMV的外壳蛋白编码基因设计了特异引物和特异Taqman探针,特异引物的扩增片段约为500bp,阳性质粒的实时荧光PCR方法的检测下限为20fg/μL,是一步法RT-PCR方法的100倍,检测时间由约8h缩短至4h。种脐灰色斑驳的大豆种子BPMV检测呈阳性,种脐黑色斑驳的大豆种子BPMV检测呈阴性。
Abstract:Bean pod mottle virus (BPMV), which can be seedborne virus and causes yield loss and quality reduction seriously to soybean is an important quarantine pest for China. For detection of BPMV in imported soybean, the specific primers and Taqman probe were designed based on the coding gene of BPMV coat protein and one step RT-PCR and one step real-time fluorescent RT-PCR methods were developed. The aplicon of one step RT-PCR was about 500bp, and the detection limit for real-time fluorescent PCR was 20fg/μL of positive cloning plasmid, while that was 100-fold higher than that of RT-PCR, and the detection duration shortened from at least 8h to 4h . The detection result of soybean seeds with gray mottled hila by one step real-time fluorescent RT-PCR showed positive, while that of soybean seeds with black mottled hila showed negative.