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进境大豆携带的病毒主要有菜豆荚斑驳病毒(BPMV)、烟草环斑病毒(TRSV)、烟草条纹病毒(TSV)和南方菜豆花叶病毒(SBMV)等,均为大豆种传病毒,可随大豆种子实现远距离传播。本研究对这4种大豆病毒的外壳蛋白基因部分序列设计引物,并通过优化引物、模板浓度和扩增参数,在一个体系中成功对4种病毒进行了多重RT-PCR扩增,得到307bp、206bp7、17bp5、18bp共4条特异性条带,建立了能同时检测BPMV、TRSV、TSV和SBMV的多重RT-PCR检测体系。该方法快速、灵敏、简便,同时特异性强,在出入境检疫中具有广泛的应用前景。
The major carriers of imported soybean are BPMV, TRSV, TSV and SBMV, all of which are soybean-borne viruses. Soybean seeds to achieve long-distance transmission. In this study, primers were designed for partial sequences of the coat protein genes of four soybean viruses. Multiple primers were designed to amplify four kinds of viruses in a system by optimizing primer, template concentration and amplification parameters. The results showed that 307bp, 206bp7, 17bp5 and 18bp. A multiplex RT-PCR detection system was established for the simultaneous detection of BPMV, TRSV, TSV and SBMV. The method is rapid, sensitive and simple, and has high specificity and wide application prospect in exit quarantine inspection.