Objective To understand the occurrence of healthcare-associated infection (HAI) in intensive care unit (ICU) through conducting targeted monitoring, and provide scientific basis for reducing the occurrence of HAI. Methods Patients in the ICU in a hospital from May 2010 to December 2022 were selected as research objects. Targeted monitoring was carried out. Incidence of HAI, incidence of device-associated infection, distribution of pathogens, sites of HAI, and compliance rate of hand hygiene were statistically analyzed. Results A total of 8 998 patients were admitted to the ICU, with a HAI rate of 10.35% and a HAI case rate of 14.66%. The adjusted HAI case rate was 3.72%. Difference in HAI rates among different years was statistically significant (P < 0.001). Utilization rates of ventilators, urinary catheters, and central venous catheters were 57.56%, 95.92%, and 79.14%, respectively. Incidences of ventilator-associated pneumonia (VAP), catheter-associated urinary tract infection (CAUTI), and central line-associated bloodstream infection (CLABSI) were 10.62‰, 3.08‰, and 0.65‰, respectively. Utilization rate of ventilators increased from 37.12% in 2010 to 72.35% in 2022, and the incidence of VAP dropped from 23.36‰ to 5.61‰. Incidences of CAUTI and CLABSI tended to be relatively stable. A total of 1 673 pathogenic bacteria strains were detected, and the top 5 pathogens were Acinetobacter baumannii (34.79%), Klebsiella pneumoniae (18.35%), Pseudomonas aeruginosa (10.58%), Stenotrophomonas maltophilia (2.81%) and Escherichia coli (2.75%). The top 3 detected multidrug-resistant organisms (MDROs) were carbapenem-resistant Acinetobac-ter baumannii (CRAB, 83.88%), methicillin-resistant Staphylococcus aureus (MRSA, 56.37%) and carbapenem-resistant Klebsiella pneumoniae (CRKP, 50.29%). MDROs with the top 3 highest HAI rates were CRAB (6.39%), CRKP (2.56%) and carbapenem-resistant Pseudomonas aeruginosa (CRPA) (1.09%). The main site of HAI was respiratory system (55.80%), followed by urinary system (16.53%), blood system (16.38%), and digestive system (6.75%). Hand hygiene compliance rate of ICU health care workers increased from 62.25% in May-December 2010 to 87.86% in 2022. The consumption of alcohol-based hand rub and hand sanitizers increased from 65.66 mL/day/bed in 2013 to 95.49 mL/day/bed in 2022. Conclusion The targeted monitoring results of HAI in ICU in the past 13 years help health care workers understand the trend of HAI, provide powerful data support for formulating effective prevention and control measures, and reduce the occurrence of HAI.