烟草环斑病毒RT-RealtimePCR检测方法

Detection of Tobacco ringspot virus by RT-Realtime PCR

烟草环斑病毒(Tobacco ringspotvirus,TRSV)是我国公布的二类检疫危险性有害生物,对农业生产危害较大。受该病毒侵染的一些寄主植物出现隐症,并伴随病毒浓度降低,给检测工作造成困难。根据TRSV外壳蛋白基因序列设计合成了一对引物及一条MGB探针,优化了反应条件,建立了TRSV实时荧光RT-PCR检测方法。该方法与常规的DAS-ELISA方法和RT-PCR方法相比,灵敏度分别提高了200倍和4倍,并且对不同寄主上的TRSV均有较好的检测效果。

Tobacco ringspot virus was dangerous quarantine pest defined by P. R. China. Some hosts infected by the virus appeared masking symptom with lower virus concentration. Therefore, it is difficult for the virus detection. In the study, according to coat protein gene sequence, a pair of primers and a MGB probe were designed. The reaction condition was optimized. As a result, the detection method of TRSV by RT-Realtime PCR was established. This detection method was 200 and 4 times more sensitive than DAS-ELISA and RT-PCR respectively, and suitable to detect TRSV from different host plants.