We investigated different types of Shiga toxin-producing Escherichia coli (STEC) not belonging to serogroup O157 for their role as human pathogens. Non-O157 STEC isolated from 89 human patients in Germany were characterized according to serotypes, virulence markers, and association with human illness. EaeA-positive STEC were isolated from 54 (60.7%) of the patients and were frequently associated with severe diarrheal disease, hemolytic uremic syndrome, and young age. EaeA-negative STEC were found in 35 (39.3%) of the patients and were more associated with clinically uncomplicated cases and adult patients. For pediatric patients, a serotype-independent diagnosis of STEC is recommended.