Streptococcus pneumoniae is the third most frequent etiologic agent of bacterial meningitis in Poland. Therefore, the objective of this study was to characterize S. pneumoniae isolates responsible for meningitis in Poland from 1997 to 2001 by determining their serotypes, antimicrobial susceptibility, and genetic relatedness. Among 36 serotypes and rough isolates, the most common were serotypes 3, 8, 19F, 6B, 4, and 23F, altogether accounting for 45.3% of cases. Penicillin-nonsusceptible pneumococci constituted 14.8% of all isolates. A high heterogeneity of pneumococcal isolates concerning serological features as well as genetic relatedness based on pulsed-field gel electrophoresis analysis was observed. With the exception of some serotypes (e.g., 3, 7F, 19F), isolates of the same serotype do not appear to be more closely related to each other than to isolates of different serotypes.