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本研究建立了大豆种子中菜豆荚斑驳病毒(BPMV)和烟草环斑病毒(TRSV)单管双重实时荧光PCR检测方法。将含有相同浓度的分别带有BPMV和TRSVCP基因的质粒溶液作为阳性对照,以受两种病毒侵染的大豆种子作为待测样品进行实时荧光PCR检测,结果表明能从同一管中同时检测出这两种病毒而不发生交叉反应。尽管在阳性对照中,二者的检测限相当,均可达到35 pg/mL,但在实际应用中,两种病毒由于在大豆种子中的浓度不一致而存在一定的差别。该方法快速、灵敏、简便,同时特异性更强,在出入境检验检疫中具有广泛的应用前景。
In this study, we established a single-tube double real-time PCR assay for detecting soybean seed pod mottle virus (BPMV) and tobacco ring spot virus (TRSV). Plasmid solutions containing the BPMV and TRSVCP genes at the same concentration as the positive control and the soybean seeds infected with both viruses as the samples to be tested were subjected to real-time fluorescence PCR. The results showed that simultaneous detection of this from the same tube Both viruses do not cross-react. Although in the positive control, the detection limit of both can reach 35 pg / mL, but in practical application, there are some differences between the two viruses due to the inconsistent concentration in soybean seed. The method is rapid, sensitive and simple, and has higher specificity at the same time, and has wide application prospect in exit-entry inspection and quarantine.