Objective To evaluate the health and economic burden from the perspective of hospitals after implementing pre-isolation based on the "Standard for infection prevention and control of carbapenem-resistant Entero-bacterales" (WS/T 826-2023). Methods Patients from 4 internal medicine intensive care units (ICUs) in 2 tertiary first-class hospitals in Urumqi were selected as research subjects. Data of patients from February 1, 2023 to February 1, 2024 (control group) and from February 2, 2024 to February 1, 2025 (intervention group) were collected retrospectively. Pre-isolation measures recommended by WS/T 826-2023 were implemented in the intervention group, while not in the control group. Compared with patients without Enterobacterales infection, the mortality risk, extra length of hospital stay, and additional hospitalization costs of patients infected with carbape-nem-sensitive Enterobacterales (CSE) and carbapenem-resistant Enterobacterales (CRE) were calculated with Cox proportional hazards regression model and multi-state model. Results A total of 5 286 ICU patients were included in the analysis, with 2 806 in the control group and 2 480 in the intervention group. The detection rate of CRE in the intervention group was lower than in the control group (2.82% vs 4.88%), and the mortality also decreased obviously in the intervention group (7.38% vs 13.44%), differences were both statistically significant (both P<0.05). In the intervention group, there was no statistically significant difference in the mortality risk between patients with CRE infection and those without Enterobacterales infection (P>0.05). The intervention group had an increase in extra length of hospital stay of 2.03 days per case and an increase in additional hospitalization cost of 11 581.35 Yuan per case due to CRE infection. Conclusion After implementing pre-isolation based on "Standard for infection prevention and control of carbapenem-resistant Enterobacterales" (WS/T 826-2023), the extra length of hospital stay and economic burden in CRE-infected patients increased. However, the detection rate of CRE and the mortality of patients also decreased.