Abstract: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.