BackgroundStreptococcus pneumoniae is a major cause of bacterial infection among infants and young children with high morbidity and mortality. The serotype distribution of S. pneumoniae varies with geography, time, age, and disease.AimWe aimed to investigate the current status of molecular characteristics of S. pneumoniae strains isolated from pediatric patients in Shanghai, China.MethodsBetween 2016 and 2018, 73 clinical S. pneumoniae isolates were characterized by capsular serotype, multilocus sequence typing, antibiotic susceptibility, and resistant genes.ResultsThe most common serotypes were 19F (39.7%), 19A (16.4%), 6A (11.0%), 14 (9.6%), and 6B (8.2%). The coverage rates of the 7‐, 10‐ and 13‐valent pneumococcal conjugate vaccines were 64.4%, 64.4%, and 91.8%, respectively. The five predominant sequence types were ST271 (37.0%), ST320 (19.2%), ST3173 (11.0%), ST876 (6.8%), and ST81 (4.1%), which were mainly associated with serotypes 19F, 19A, 6A, 14, and 23F, respectively. The rates of resistance to penicillin and ceftriaxone were 21.9% and 39.7%, respectively. All strains displayed resistance to macrolides, 54.8% of which possessed both erm(B) and mef(A/E) genes, and 41.1% carried the erm(B) gene alone. Tn2010 (41.1%) was the most common transposon.ConclusionsClonal complex 271 (Taiwan19F‐14 clone) played a dominant role in the dissemination of pneumococcal isolates. The prevalent serotypes indicated a lack of the 7‐valent pneumococcal conjugate vaccine, which has not been included in national immunization programs in mainland China. The high rate of macrolide resistance made the empirical use of macrolides alone not suitable for treating pediatric pneumococcal disease.