Objective To study carbapenem-resistant Enterobacterales (CRE) infection in the neurosurgical intensive care unit (NS-ICU) under the combination of active screening using anal swabs and bundled intervention mea-sures in environment with high prevalence of CRE infection, and provide scientific basis for healthcare-associated infection (HAI) prevention and control. Methods Inpatients in NS-ICU from 2020 to 2021 were selected as the control group, and didn't undergo active anal swab screening. NS-ICU inpatients from 2022 to 2023 were taken as the trial group, and implemented intervention of active anal swab screening. Isolation measures for CRE positive cases in two groups were implemented, and the incidence of CRE HAI between two groups of patients were compared. Results 645 patients were actively screened for CRE, with a total of 912 screenings. The overall screening positive rate was 9.21%, and the rectal CRE colonization rate in NS-ICU patients was 13.02% (84/645), out of which Klebsiella pneumoniae accounted for 92.86% (n=78 strains). The positive screening rates in patients in the trial group within 48 hours after admission as well as on day 3-7, day 8-15, and 16-30 were 3.30%, 4.94%, 13.68%, and 18.85%, respectively. With prolonged hospitalization time, the positive rate of CRE anal swab screening gradually increased. During the study period, the overall CRE HAI rate was 6.24% (72/1 153). After implementing active screening intervention, the CRE HAI rate in the trial group was lower than that in the control group (3.57% [23/645] vs 9.65% [49/508], P < 0.001). Conclusion In hospital environment with high CRE prevalence, implementing active screening and bundled intervention measures can effectively identify CRE carriers and reduce the risk of infection caused by intestinal colonization.