Objective To investigate the pathogen distribution of infection in children with pediatric intensive care unit (PICU) and analyze the drug resistance so as to provide guidance for clinical prevention and treatment. Methods From Jun. 2014 to Dec. 2016, the infection data of 5754 children with PICU in our hospital were analyzed retrospectively. Pathogenic bacteria identification and drug sensitivity test were carried out to analyze the distribution of pathogens and drug resistance. Results The incidence rate of infection in PICU was 14.11% (812/5754) which was significantly higher than that in the general pediatric ward 7.68% (1808/23546), the difference was statistically significant (χ2 =235.039, P<0.05), among the 812 strains of pathogenic bacteria, gram negative bacilli (G- ) were the main pathogens (59.24%), and the pathogen bacteria were mainly distributed in the lower respiratory tract (63.66%), urinary tract (17.12%) and blood system (11.95%). G- whose resistance rate to piperacillin was 100%, and resistancerates to aztreonam and other cephalosporins were above 50%, was mainly Klebsiella pneumoniae. G+ (mainly Enterococcus faecium) whose resistance rates to penicillin, sulfamethoxazole, clindamycin and ampicillin were 100%, were mainly distributed in the urinary tract. G+ had good sensitivity to linezolid and linezolid, but vancomycin resistant strains were also appeared. Conclusion The incidence of PICU infection is high, and the situation of drug resistance is severe. Therefore, proper antibiotics should be applied to prevent the spread of nosocomial infection.