ObjectiveTo investigate the pathogens causing healthcareassociated infection (HAI) in a hospital, and guide the prevention and control of HAI.MethodsDistribution and antimicrobial resistance of pathogens isolated from patients with HAI in 2012 were analyzed statistically.ResultsA total of 793 isolates of 71 pathogen species were isolated, including 504 isolates (63.56%) of gramnegative bacteria，204 (25.72%）of grampositive bacteria, and 85 (10.72%） of fungi; the top five pathogens were Escherichia coli（n=129，16.27%）, Klebsiella pneumoniae（n=121，15.26%）, Acinetobacter baumannii（n=100，12.61%）, Staphylococcus aureus（n=79，9.96%）, and Pseudomonas aeruginosa（n=70，8.83%）， the top three sites of pathogen islation were lower respiratory tract (323,40.73%), blood stream (116.14.63%)， and urinary system (116,14.63%).274(34.55%) pathogens were multidrugresistant organisms.ConclusionThe major pathogens in HAI are gramnegative bacteria, most are multidrugresistant strains, monitor should be intensified.