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Objective: The rising number of carbapenemase-resistant Enterobacteriaceae (CRE) cases has become a concern worldwidely. This study investigated patient characteristics with CRE and analyzed the risk factors associated with its acquisition. Methods: A retrospective review of the electronic medical records of the Kangbuk Samsung Medical Center from May 2016 to April 2019 was performed. The inclusion criterion was hospitalized patients aged ≥18 years with confirmed CRE acquisition. Patients were divided by CRE acquired and non-required patients. CRE acquired patients were those with CRE confirmed by their active surveillance cultures, while non-acquired patients were those with carbapenemase-sensitive Enterobacteriaceae (CSE). If CRE was isolated more than once during hospitalization, only the first isolation was used for data analysis. Patient characteristics, antibiotic used, and the duration of use were compared between two groups using univariate analysis, and the risk factors associated with CRE were analyzed using multiple logistic regression analysis. Results: Among the 73 CRE acquired patients, 44 (60.3%) were positive for carbapenemase-producing Enterobacteriaceae (CPE). Infection from Klebsiella pneumonia (42 cases, 57.5%), Escherichia coli (17 cases, 23.3%), and Enterobacter cloacae (5 cases, 6.8%). The risk of CRE acquisition was significantly increased by 4.99 times [confidence interval (CI), 1.40-17.78; p=0.013] with mechanical ventilation, 3.86 times (CI, 1.59-9.36; p=0.003) with penicillin administration, and 21.19 times (CI, 6.53-68.70; p<0.001) with carbapenem administration. Conclusions: Proper antibiotic use including the selection, frequency, and duration, and patients on mechanical ventilators need close monitoring.