ObjectiveTo understand the clinical distribution characteristics and antimicrobial resistance of pathogens causing healthcareassociated infection(HAI) in a comprehensive hospital. MethodsClinical data of patients with HAI in this hospital between  May 2012 and May 2015 were collected, the distribution and antimicrobial resistance of pathogens isolated from patients were analyzed.ResultsA total of 6 563 cases of HAI occurred among 183 850 patients, incidence of HAI was 3.57%, 445 patients were isolated at least two kinds of pathogens, 375（84.27%） patients were isolated two kinds of pathogens, 132 of whom were infected with both gramnegative bacilli. 4 478 specimens were sent for pathogenic detection, 2 503 (55.90%) of which were isolated pathogens; a total of 2 755 pathogens were isolated, including 1 713(62.18%) strains of gramnegative bacilli, 732(26.57%) grampositive cocci, 304(11.03%) yeastlike fungi, and 6(0.22%) anaerobic bacteria. 524(19.02%）strains were mainly from patients in department of neurology. The main specimen was sputum (n=1 340, 48.64%). The isolation rates of carbapenemresistant Escherichia coli(CREC), Klebsiella pneumoniae (CRKP), Acinetobacter baumannii (CRAB), and Pseudomonas aeruginosa(CRPA) were 0.39%（2/510）, 1.66%（3/181）, 59.14%（207/350）, and 5.29%（11/208）respectively；isolation rate of methicillinresistant Staphylococcus aureus (MRSA) was 21.55%（25/116）. ConclusionMultidrugresistant organisms causing HAI are various, it is necessary to understand distribution characteristics and prevalence of pathogens, monitor multidrugresistant organisms, and implement contact isolation measures, so as to prevent the outbreak of HAI.