Objective To investigate healthcare-associated infection(HAI)in different departments in a children’s hos-pital,and provide scientific basis for effective prevention and control of HAI in children.Methods By bed-side visiting and medical record reviewing,survey on HAI prevalence rate was conducted among hospitalized patients in a given day in 2012, 2013,and 2014 respectively,and surveyed data of three years were analyzed.Results HAI prevalence rates in 2012,2013, and 2014 were 6.52%,5.59%,and 5.85% respectively,there was no significant difference (χ2 =1.02,P =0.60);the average HAI prevalence rate in three years was 5.98%,there was significant difference in average HAI prevalence rate among different departments(χ2 =291.83,P <0.001 ),the top 4 departments of HAI prevalence rate was hematology (27.49%),cardiothoracic surgery (21.13%),neonatal intensive care unit (16.43%),and pediatric intensive care unit (12.76%);the main infection sites were lower respiratory tract (58.44%),upper respiratory tract (22.63%),and gas-trointestinal tract (6.58%);the main pathogens causing HAI was Klebsiella pneumoniae (n=32,28.32%),followed by Acinetobacter baumannii (n=9,7.96%),Escherichia coli (n=9,7.96%),and Pseudomonas aeruginosa (n=7,6.19%). Conclusion Strengthening prevention and control intervention of HAI in high risk departments has important significance to reduce the incidence of HAI in children’s hospital.