Nasopharyngeal carriage is a major factor in the transmission of pneumococcal disease. The aim of this study was to determine the prevalence of asymptomatic nasopharyngeal carriage of Streptococcus pneumoniae and the distribution of serogroups and serotypes in children aged 3-36 months attending day care centres in Belgium. A single nasopharyngeal swab was cultured from 467 children attending 30 different day care centres between December 2000 and March 2001. S. pneumoniae isolates were serotyped and their antibiotic susceptibilities assessed by disk diffusion. The overall nasopharyngeal carriage rate for S. pneumoniae was 21% in the 467 children. None of the commonly accepted risk factors studied was associated significantly with carriage. Capsular serotypes isolated were 19F (27.3%), 6B (20.2%), 23F (19.2%), 19A (10.1%), 6A (7.1%), 14 (5.1%) and others (11.0%). Theoretical coverage by the seven-valent (serotypes 4, 6B, 9V, 14, 18C, 19F and 23F) pneumococcal conjugate vaccine was 73.7%. Fourteen (14.1%) of 99 strains were non-susceptible to penicillin, 48 (48.5%) to tetracycline and 61 (61.6%) to erythromycin. Theoretical coverage by the seven-valent pneumococcal conjugate vaccine was 93% for the penicillin-resistant serotypes, 69% for the tetracycline-resistant serotypes and 75% for the erythromycin-resistant serotypes. Use of the seven-valent pneumococcal conjugate vaccine could potentially reduce nasopharyngeal carriage of the antibiotic-resistant strains.