Abstract:The total 73 different tobacco virus disease samples collected from different regions in Yunnan were detected with electron microscopy, DAS-ELISA, immunostrip test and multiplex RT-PCR. The results showed that the tobacco virus diseases were serious in Yunnan Province. 10 kinds of viruses were detected in the experiments. The viruses were more at the northeast and central part, such as Chuxiong and Wenshan, than the north part of Yunnan. The results showed that main viruses were Tobacco mosaic virus(TMV), Cucumber mosaic virus (CMV), Potato virus Y(PVY), Tobacco etch virus (TEV) and Tobacco vein banding mosaic virus(TVBMV). The detection results showed there were nine types complex infections. The complex infection sample was 31.50% among the total testing samples. The main type of complex infection was that TEV and TVBMV, occupied 34.78% among the all combination.