Abstract: OBJECTIVE To analyze the drug susceptibility of 224 strains of Enterobacteriaceae to 21 kinds of antibiotics in the hospital,so as to guide rational use of antibiotics. METHODS A total of 224 strains of Enterobacteriaceae infection specimens from Aug.2014 to Apr.2016 in the hospital were retrospectively analyzed. The distribution of Enterobacteriaceae in different specimens and departments were analyzed, the bacteria were identified and their drug susceptibility were analyzed by the susceptibility analyzer and the susceptibility of Enterobacteriaceae to 21 kinds of antibiotics were detected. RESULTS Among 224 strains of Enterobacteriaceae isolates, 42.86%(96 strains) were isolated from the sputum, 27.68%(62 strains) from the urine, and 12.05%(27 strains) from the blood; The top three departments were internal medicine(68.30%, 153 strains), surgery(12.50%, 28 strains) and outpatients(11.16%, 25 strains), respectively. The drug resistant rates of 224 strains of Enterobacteriaceae to piperacillin/tazobactam, cefoperazone/sulbactam, and imipenem were 1.3%, 1.8%, and 4%. respectively , The drug resistant rate to ampicillin was the highest, which was 84.4%. The drug resistant rates to lomefloxacin, levofloxacin, and ciprofloxacin were 47.8%, 44.2%, and 46%, respectively. CONCLUSION Enterobacteriaceae are mainly isolated from respiratory system and urinary system, the drug resistant rates were low to carbapenems,β-lactamase/β-lactamase inhibitor, while the drug resistant rates of Enterobacteriaceae isolated from urine to quinolone were higher than from other specimens. The issue should be paid more attention and we should strengthen the rational use of antibiotics.