ObjectiveTo investigate the distribution and antimicrobial resistance of pathogenic bacteria in a pediatric intensive care unit (PICU), and provide reference for preventing and controlling healthcareassociated infection and rational use of antimicrobial agents. MethodsIsolation rate and distribution of pathogenic bacteria isolated from PICU patients from June 2010 to May 2011were analyzed statistically. Results A total of 245 pathogenic bacterial strains were isolated, 73.88%, 22.86% and 3.26% were gramnegative bacteria, grampositive bacteria and fungi, respectively. The isolation rate of Klebsiella pneumoniae, Escherichia coli, Pseudomonas aeruginosa, Acinetobacter baumannii, Streptococcus pneumoniae and Staphylococcus aureus was 18.78%, 14.29%, 11.43%, 8.16%, 5.31% and 4.90%, respectively. The percentage of extendedspectrum βlactamaseproducing Klebsiella pneumoniae and Escherichia coli was 11.63% and 8.85%, respectively; detection rate of methicillinresistant Staphylococcus aureus was 16.67%. Most gramnegative bacteria were sensitive to carbapenems and cefoperazone / sulbactam, and relatively resistant to other antimicrobial agents. Acinetobacter baumannii was highly resistant to carbapenems(75.00%). Streptococcus pneumoniae and Staphylococcus aureus were highly sensitive to ceftriaxone, cefazolin and cefepime(0.00%-7.69%); no bacterial strain was found to be resistant to vancomycin. ConclusionGramnegative bacteria is the main pathogenic bacteria in PICU patients, most isolates are highly drugresistant. Analysis of pathogenic bacteria and drugresistance surveillance are of vital importance to guide treatments for critically ill children and reduce drugresistant bacterial strains.