ObjectiveTo explore the current status of  healthcareassociated infection (HAI) and antimicrobial use in a children’s hospital.  MethodsPrevalence rates of HAI and antimicrobial use among hospitalized patients at 0∶00—24∶00 of May 1, 2014 were investigated by combination of bedside visiting and medical record reviewing. ResultsA total of 1 027 patients were investigated, 8 patients developed 10 times of infection, prevalence rate of HAI was 0.78%, prevalence case rate was 0.97%. HAI mainly occurred in patients in blood center (n=4),  the main infection site was respiratory tract（upper respiratory tract, n=2; lower respiratory tract，n=2）, antimicrobial usage rate was 62.12%. Antimicrobial usage rate， purpose of antimicrobial use， and combination use of antimicrobial agents among different departments were all significantly different(all P<0.05).The departments with top 3 antimicrobial usage rates were neonatal center（89.69%）, emergency center（76.00%）, and comprehensive department（73.91%）；except department of ophthalmologyotorhinolaryngologystomatology (preventive antimicrobial use accounted for 57.89%) and department of surgery(therapeutic antimicrobial use accounted for 26.32%), the other departments mainly used therapeutic antimicrobial agents; department of ophthalmologyotorhinolaryngologystomatology, heart center, and neurological rehabilitation center mainly adopted single medication treatment （all ＞95%），twodrug combination rate in neonatal center accounted for 48.28%, threedrug combination rate in blood center accounted for  30.30%. ConclusionRoutine surveillance on departments and sites of high HAI incidence should be intensified in children’s hospitals, training on knowledge of HAI among health care workers should be strengthened， and antimicrobial should be used rationally.