Objective To understand the current situation and changing trend of healthcare-associated infection(HAI) in a hospital of Qinghai Province from 2010 to 2017. Methods Medical record review and bedside survey was adopted to investigate the occurrence of HAI, antimicrobial use, pathogens causing HAI, and bacterial culture(therapeutic antimicrobial use) of all inpatients in one day of each year from 2010 to 2017. Results A total of 12 446 inpatients were investigated in 8 cross-sectional surveys, 488 patients had 555 cases of HAI. HAI rate and HAI case rate were 3.92% and 4.46% respectively. Prevalence of HAI in each year were 2.93%-5.70%. The highest prevalence of HAI was found in ICU (58.72%), and the main site of HAI was lower respiratory tract (n=279). There was a significant difference in the usage rate of antimicrobial agents in each year (χ²=135.98,P<0.01), decreased from 55.56% in 2010 to 16.05% in 2017. The use of antimicrobial agents were mainly therapeutic (65.74%), proportion of therapeutic antimicrobial use gradually declined; 91.52% were antimicrobial monotherapy, proportion of combined use of at least three antimicrobial agents was 0 in 2011-2017. There was a significant difference in pathogenic detection of therapeutic antimicrobial use among each year (χ²=57.48,P<0.05). A total of 203 pathogens were isolated, the main pathogens were gram-negative bacteria (n=138, 67.98%) and gram-positive bacteria (n=52, 25.62%), followed by fungi and other pathogens. Conclusion The prevalence of HAI in this hospital is generally stable, antimicrobial use and pathogenic detection of therapeutic antimicrobial use are gradually standardized, close attention should be paid to the management of HAI in ICU, departments of neurosurgery and hematology.