Abstract: Objective To investigate the nosocomial infection prevalence in a tertiary hospital, so as to provide the scientific basis for the prevention and control of nosocomial infection. Methods A cross-sectional survey method was used to investigate the nosocomial infection prevalence of five consecutive years (2010-2014). Surveys were conducted on one day each year in November according to the standards made by Anhui Center for Quality Control of Nosocomial Infection. Results In 2010-2014, on the annual survey day, the nosocomial infection rate of hospitalized patients was 3.19%, 2.32%, 3.03%, 2.34% and 1.74%, respectively. The lower respiratory tract infection site was in the first place of each year, followed by urinary tract, and upper respiratory tract infection. 91 strains of pathogen and microorganisms were isolated from the cases with hospital infection. The five leading pathogeny microorganism were as following: escherichia coli (19 stains), pseudomonas aeruginosa(15 stains),acinetobacter baumannii (12 stains),fungus (9 stains),staphylococcus aureus(5 stains). For 5 consecutive years, the antibiotic used rates were 48.88%, 40.04%, 37.59%, 36.85% and 36.23%, respectively.Gram negative bacilli were the dominative pathogenic bacteria for nosoeomial infection. Conclusions Consecutive five-year investigation of the hospital inpatient hospital infection rates is in the standard range. The use of anti-biotics of control rates is in the standard range. The department with high incidence of nosocomial infection department should be taken as the target of control management.