Abstract: The aim of the present study was to detect the mixed infection of the tobamoviruses (TMV and ToMV) simultaneously. The primers were designed for the movement protein regions of TMV and ToMV and synthesised. The primers were used to detect the TMV and ToMV in infected leaves. To confirm the results of molecular detection, Indirect Enzyme Linked Immunosorbent Assay (Indirect ELISA) was used to detect the viruses. Based on the results of the present study, the movement protein gene may be used as candidate gene for the identification of plant viruses in plant pathology laboratories. Mixed infection of both viruses to same host can be detected by using duplex RT-PCR method. The assay could be performed in a single tube for simultaneous detection of TMV and ToMV.