Nine cases of midline T-cell lymphoma (MTL) were studied for expression of Epstein-Barr virus (EBV) encoded small RNA-1 (EBER-1) and EBV latent membrane protein-1 (LMP-1) using in situ hybridization and immunohistochemistry respectively. EBER-1 was detected in nuclei of tumor cells in 8/9 cases of MTL and LMP-1 was found on membrane and in cytoplasm of tumor cells in 7/9 cases of MTL. The results show: (1) There is a strong association between EBV and MTL. EBV may play an important role in the pathogenesis of MTL. (2) The detection frequency of EBV in MTL is higher than peripheral T-cell lymphoma from other sites of the body. (3) EBER-1 in situ hybridization and LMP-1 immunohistochemistry are sensitive and reliable in detection of EBV in MTL and the latter is more economic and convenient.