This study examined the epidemiology, antibiotic susceptibility, and serotype distribution of Streptococcus pneumoniae associated with invasive and noninvasive pneumococcal disease in children in China. A total of 451 clinical isolates from children (age, <5 years) were collected from 8 cities from January 2005 to December 2006, including 31 isolated from invasive disease. In vitro susceptibility to 14 antimicrobial agents was determined by the agar dilution method. Among all isolates tested, 64.3% were resistant to penicillin, and for invasive isolates, the resistance rate was 55.2%. Isolates from Wuhan and Nanjing showed the highest prevalence of penicillin resistance (89.6% and 85%), followed by those from Shenzhen (72.4%) and Chengdu (56.7%). Multidrug resistance rates to tetracycline, erythromycin, and clindamycin reached 90%. The 6 most common serotypes were 19F, 19A, 14, 6B, 23F, and 15. These accounted for 80.7% of the isolates. Resistance to penicillin varied among the 6 leading serotypes, ranging from 20% in serotype 15 to 87.3% in serotype 19F. Each of the 8 cities had different serotype distribution. The potential coverage by 7-, 10-, and 13-valent pneumococcal conjugate vaccine were 63.6%, 64.8%, and 79.6%, respectively.