Symptoms associated with eggplant little leaf and leaf curl disease of eggplant was observed in the fields of Meerut district of Western Uttar Pradesh, India. Total DNA extracted from symptomatic and asymptomatic plants was subjected to nested-PCR assay using phytoplasma 16S rDNA primers and generic PCR with universal begomovirus coat-protein (CP) gene primers. Amplicons of expected size were obtained from symptomatic plants in both PCR assays. No PCR amplifications were observed for asymptomatic plants. A multiplex nested PCR assay was also developed by optimizing reaction components and reaction cycling parameters for simultaneous detection of Tomato leaf curl virus (ToLCV) and a phytoplasma. The eggplant infecting phytoplasma was identified as a member of group 16SrVI, ‘‘Candidatus Phytoplasma trifolii’’, while the begomovirus showed 99% of CP gene identity with other whitefly transmitted begomoviruses including Solanum nigrum and Tomato leaf curl virus (ToLCV). Results indicate that both phytoplasma and begomovirus can co-infect the same host plant and the multiplex PCR provides a sensitive, rapid and low cost method for simultaneous detection of little leaf of eggplant and whitefly –transmitted Tomato leaf curl virus.