ObjectiveTo investigate the distribution and antimicrobial resistance of pathogens isolated from blood culture of children in a pediatric intensive care unit (PICU), provide reference for empirical treatment of bloodstream infection in critically ill children.MethodsPathogenic bacteria isolated from blood culture of children in a PICU in 2011-2015 were identified and performed antimicrobial susceptibility testing.ResultsA total of 180 strains of pathogens were isolated from 3 215 blood specimens, the positive rate was 5.60%, 153（85.00%） of which were grampositive bacteria and 27（15.00%） were gramnegative bacteria. The top five isolated pathogens were Staphylococcus epidermidis(26.67%)， Staphylococcus hominis(25.00%)，Staphylococcus haemolyticus(11.66%)， Escherichia coli(5.55%)，and Staphylococcus aureus(3.89%). The resistance rates of Staphylococcus spp. to linezolid, vancomycin, and quinupristin/dalfopristin were all 0; the detection rates of methicillinresistant coagulasenegative staphylococci (MRCNS) and methicillinresistant Staphylococcus aureus(MRSA) were 70.18% and 42.68% respectively； Escherichia coli had high resistance rates to ampicillin， cefazolin， ceftriaxone， gentamycin, and compound sulfamethoxazole（50.00%-80.00％）.ConclusionCNS and Escherichia coli are the main pathogens in blood culture of children in PICU, differences in antimicrobial resistance exist among different types of CNS.