BACKGROUND: Viral isolation in cell culture remains as a reference method for diagnosis of enteroviral infection. Enteric adenoviruses are cultivated in 293 cells. Enteroviral and enteroadenovrial tropisms for the gastrointestinal tract lead to the assumption that 293 cells would be useful in enteroviral isolation. We evaluated usefulness of 293 cells in the diagnosis of enteroviral infection. METHODS: Human embryonic lung fibroblasts (HEL), HeLa, RD and 293 cells were used to evaluate viral isolation from clinical specimens, susceptibilities of the cell lines to reference enteroviral strains and influence to stool extracts on the viral isolation. Forty-four stool specimens collected from patients during the epidemic period of type 9 echoviral aseptic meningitis and type 30 echoviral culture-positive 33 stool and 58 cerebrospinal fluid (CSF) specimens were inoculated onto cell lines. RESULTS: Echovirus type 9 was isolated from 31 of 44 stool specimens. Of 31 echovirus 9 isolates, 22 (71.0%), 21 (67.7%), 6 (19.4%) and 3 (9.7%) were detected in HEL, 293, RD and HeLa, respectively. Of 33 echovirus 30 isolates from stool specimens, 32 (97.0%) were detected in 293; 17 (51.5%) were detected in RD. Of 58 echovirus 30 isolates from CSF specimens, 39 (67.2%) were detected in 293; 30 (51.7%) were detected in RD. 293 cells were sensitive for coxsackievirus A9 reference strain and echovirus 7 reference strain. Stool extracts induced enhanced cytopathic effect by echovirus 9 infection in 293 and HEL. CONCLUSIONS: 293 cells are useful in the diagnosis of echoviral and some enteroviral infection.