ObjectiveTo investigate the prevalence rate of healthcareassociated infection(HAI) in a hospital, and evaluate the reliability and accuracy of routine HAI surveillance data. MethodsHAI prevalence rate survey was conducted among all inpatients from 0∶00 to 24∶00 on investigation day with crosssectional survey, uniform questionnaires were filled out,and data were analyzed.ResultsOn June 9, 2010 and April 8, 2011, 1 196 inpatients were investigated, the actual investigation rate was 97.04% and 97.58% respectively; HAI prevalence rate was 6.27% and 6.11% respectively, which was slightly higher than HAI of the respective month (5.01% and 5.54% respectively). The main infection site was lower respiratory tract(66.27%), followed by urinary tract(13.25%) and upper respiratory tract(7.23%). ConclusionPrevalence survey can benefit to realize the current status of HAI, improve routine surveillance quality, and take proper measures to reduce HAI rate through the intervention in departments with high HAI.