The objectives were to study serotypes and antibiotic susceptibility of Streptococcus pneumoniae carried by healthy children attending a day-care centre in St. Petersburg. S. pneumoniae colonization was investigated in 125 children aged 16-70 months. Antibiotic susceptibility was determined by E-test and disk diffusion. 83 S. pneumoniae cases were isolated in 75/125 (60%) children: 36/75 (48%) in the nasopharynx, 12/75 (16%) in the oropharynx and 27/75 (36%) in both. Carriage rates were 100%, 68%, 72%, 46% and 54% in children aged 12-23, 24-35, 36-47, 48-59 and ≥60 months, respectively. 97.6% of isolates were susceptible to penicillin. 61.4%, 32.5%, 19.3%, 16.7% and 6% isolates were non-susceptible to trimethoprim/sulfamethoxazole, tetracycline, clindamycin, erythromycin and chloramphenicol, respectively. 20.5% of isolates were multidrug resistant (MDR). 45% of isolates were of serotypes included in the 7-valent pneumococcal conjugate vaccine (7V-PCV); 64.9%, 56.8%, 32.4% and 27% of 7V-PCV serotypes were resistant to trimethoprim/sulfamethoxazole, tetracycline, clindamycin and erythromycin, respectively. The respective figures for MDR isolates were 100%, 94.1%, 70.6% and 76.5%; 76.5% of all MDR isolates were covered by 7V-PCV. In conclusion: 1) resistance to trimethoprim/sulfamethoxazole and tetracycline was high; 2) resistance to macrolides was higher than in other Russian regions; 3) 7V-PCV coverage was modest, but the vaccine may potentially reduce MDR-S. pneumoniae.