Abstract: Objective To explore the common pathogens distribution and the drug resistance pattern in vitro from cerebrospinal fluid culture in children. Methods The results of cerebrospinal fluid culture in hospitalized children from January 2007 to December 2014 were retrospectively analyzed. Bacteria identification and antimicrobial susceptibility were assayed by Vitek system from Bio Mrieux Company. Some bacterial strains were tested by disk diffusion method. Results Cerebrospinal fluid from 23099 patients were tested at least one time during research period. 671 strains of bacteria were isolated with positive rate of 2.9%, among which there were 579 strains of gram positive bacteria (86.3%) and 92 strains of gram negative bacteria (13.7%). The top five bacteria were coagulase negative Staphylococcus (399 strains, 58.9%), Micrococcus (37 strains, 5.5%), Streptococcus pneumoniae (34 strains, 5.1%), Escherichia coli (32 strains, 4.8%), and Enterococcus faecium (26 strains, 3.9%). The positive rates of cerebrospinal fluid culture in 8 consecutive years showed a downward trend (χ2 = 10.410, P = 0.001). The positive rates of coagulase negative Staphylococci showed annual decline trend (χ2 = 31.200, P < 0.001). However, annual positive rates of Escherichia coli showed an upward trend (χ2 = 4.786, P = 0.029). The resistant rate of coagulase negative Staphylococcus to oxacillin was 79.8%, and no coagulase negative Staphylococcus resistant to linezolid and vancomycin was found. Micrococcus sensitive to vancomycin was 100%. The resistant rate of Streptococcus pneumoniae to penicillin was 77.8%, while the sensitive rate of Streptococcus pneumoniae to vancomycin was 100%. The sensitive rate of Escherichia coli to piperacillin/tazobactam and meropenemwas 100%. Conclusions The positive rate of cerebrospinal fluid culture is low. The main strains are coagulase negative Staphylococcus, Micrococcus, Streptococcus pneumoniae, Escherichia coli, and Enterococcus faecium. The antibiotics should been rationally chosen according to the results of bacterial strain culture and drug sensitive test.