Abstract: OBJECTIVE To discuss the detection characteristics of Enterobacteriaceae bacteria and the detection of drug resistance genes in a third-class hospital in Tianjin between 2017 to 2019. METHODS A total of 2 526 strains of Enterobacteriaceae were isolated in Tianjin People’s Hospital from Jan 2017 to Dec 2019, including 178 strains of Carbapenem-resistant Enterobacteriaceae（CRE）. The distribution characteristics and drug resistance of Enterobacteriaceae bacteria were analyzed by using an automatic microbial identification system and drug susceptibility experiments. Polymerase chain amplification reaction（PCR） was used to detect common drug resistance genes VIM, NDM, KPC, and OXA-48 in CRE bacteria. RESULTS The detection rates of CRE in Enterobacteriaceae from 2017 to 2019 were 7.52%, 6.37%, and 7.21% respectively. The strain with the highest detection rate of Enterobacteriaceae bacteria in the past three years was Escherichia coli, which was highly detected in geriatrics department, intensive care unit and respiratory medicine department. Specimens such as putum, urine, blood, alveolar lavage fluid and cerebrospinal fluid were the main sources of Enterobacteriaceae detected. The isolated E. coli was highly resistant to cefazolin, ampicillin, piperacillin, cefuroxime and ceftriaxone. Of the 178 isolated CRE strains, 168 strains were all detected to carry the carbapenase gene; a total of 146 strains were detected KPC gene, 22 strains were detected NDM gene, and no OXA-48 and VIM genes were detected. CONCLUSION The drug resistance of Enterobacteriaceae isolated in this hospital was severe, and the strains of bacteria detected were mainly E. coli and Klebsiella pneumoniae carrying the KPC gene.