Objective To analyze the characteristics and influencing factors of healthcare-associated bloodstream infection (HA-BSI) of carbapenem-resistant Enterobacterales (CRE). Methods Retrospective nested case-control study was adopted. Fifty-six patients with CRE HA-BSI in a tertiary general hospital from January 2020 to December 2022 were selected as the CRE group. With a 1:1 ratio, 56 patients with carbapenem-sensitive Enterobacterales (CSE) BSI during the same period was selected as the CSE group. Distribution of infection strains and departments was analyzed, and the relevant factors for CRE BSI were analyzed by univariate and multivariate logistic regression analyses. Results The distribution of CRE BSI was mainly in intensive care unit (ICU, n=23, 41.07%) and department of hematology (n=17, 30.36%). The main infection strains were Klebsiella pneumoniae (n=32, 57.14%) and Escherichia coli (n=16, 28.57%). Univariate analysis showed that malignant tumor, hospitalization history within 60 days, stay in ICU for >48 hours before infection, mechanical ventilation, indwelling central venous cathe-ter, combined use of at least two kinds of antimicrobial agents, and duration of antimicrobial use ≥10 days were all related to CRE BSI (all P < 0.05). Multivariate logistic regression analysis found that stay in ICU>48 hours before infection and duration of antimicrobial use ≥10 days before infection were independent risk factors for CRE HA-BSI (P < 0.05). Conclusion Clinical departments, especially ICU, should pay attention to the epidemiological history of patients, identify patients with high-risk factors for CRE BSI as early as possible, use antimicrobial agents ratio-nally and standardize invasive procedure, so as to reduce the occurrence of CRE HA-BSI.