BACKGROUND: Carbapenem-resistant Enterobacteriaceae (CRE) has increasingly become a global health problem, and the drug resistance and epidemiology of its infection are important for disease prevention and control. METHODS: The CRE isolates detected in the hospital from July 2021 to April 2023 were collected, then the antibiotic sensitivity was detected by an automatic microbial analyzer. Polymerase chain reaction (PCR) was performed to detect carbapenem-resistance genes. The homology and epidemiological characteristics of the strains of Enterobacteriaceae were analyzed by repeated common sequence PCR(ERIC-PCR). RESULTS: A total of 94 non-repeating CRE isolates were collected mainly from the respiratory tract (67.02%) and urinary tract (22.34%). Among these, 73 strains (77.66%) were carbapenem-resistant Klebsiella pneumoniae (CRKP), while 21 strains (22.34%) were carbapenem-resistant Escherichia coli (CREC). Antibiotic susceptibility testing revealed a low resistance rate to tigecycline (2.13%). Analysis of CRE resistance genes demonstrated that blaKPC accounted for 93.15% of CRKP isolates (68/73), while blaNDM was present in 90.48% of CREC isolates (19/21). In the homology analysis, CRKP isolates were divided into five clusters with C type representing the majority at 80.82% prevalence (59/73). Similarly, CREC could be classified into four clusters with A type being predominant at 80.95% among the isolated strains (17/21). The detection of CRE strains involved multiple departments in two separate wards of this hospital, located approximately 28 km from each other. CONCLUSIONS: CRE infection in this hospital has high homology and wide distribution; it is necessary to strengthen prevention and control measures to control CRE nosocomial infection spread.