Abstract: Objective To study the characteristics of S. pneumoniae isolates from respiratory specimens of children under5 years old in this region, and to provide reasonable basis for clinical treatment and epidemiology. Methods The specimens of respiratory tract were collected from 5 332 children under 5 years old with respiratory tract infection from December 2013 to November 2017, germiculture and bacteria identified were done. Multiple PCR and E-test were respectively used for serotype and drug sensitivity of the isolates, and the results were statistically analyzed. Results Two hundred and twelve S. pneumoniae isolates were isolated 5 332 children under 5 years old with respiratory tract infection, the detection rate was 3.98%. The detection rates of Streptococcus pneumoniae in male and female children were 4.19% and 3.74%, respectively. There was no statistical difference (P > 0.05). S. pneumoniae isolates detection rates were different in different seasons, the highest 5.7% in winter, and S. pneumoniae detection rates (6.7%) in winter and spring was higher than that in summer and autumn (2.0%), and the difference was statistically significant (P < 0.05). Nineteen different serotypes were identified in 212 isolates by multi-PCR, of these, 19 F accounts for up to 37.3%, followed by 19A (24.5%). S. pneumoniae are all sensitive to vancomycin and moxifloxacin. Chloramphenicol, levofloxacin, cefotaxime, amoxicillin, penicillin G, clindamycin, tetracycline and erythromycin resistant rate were 4.2%, 4.7%, 9.0%, 25.0%, 30.2%, 56.1%, 80.7% and 86.8%, respectively. Conclusion Winter is the season of the highest rate of S. pneumoniae in children under 5 years old in the region, it is necessary to perform serotype detection and drug susceptibility test so as to choice preventive vaccination and treatment antimicrobial agents.