During the year 2000 in the Rhône-Alpes region of France, 559 cases of aseptic meningitis due to enterovirus infection were recorded (approximate incidence, 10 cases per 100,000 population compared with a mean of 0.3 cases during endemic years in this region; P<0.001). Cerebrospinal fluid samples were collected from all 559 patients and processed for enterovirus RNA detection using a reverse transcriptase polymerase chain reaction assay only. In addition to the cerebrospinal fluid samples, 40 stool and 76 throat samples were collected from 116 patients (20.7% of cases); the following three enterovirus serotypes were isolated from the stool and throat samples only: ECHOvirus 13 (48/116; 41.3%), ECHOvirus 30 (44/116;37.9%) and ECHOvirus 6 (24/116, 20.7%). This is the first report that demonstrates the involvement of three different ECHOvirus serotypes, particularly ECHOvirus 13, in an outbreak of aseptic meningitis, in the French Rhône-Alpes region.