Objective To understand the targeted surveillance and risk factors for healthcare-associated infection (HAI) in a newly-built general intensive care unit (ICU), and provide basis for formulating relevant prevention and control measures of HAI. Methods Targeted surveillance on HAI in patients who were admitted to a newly-built general ICU in 2017-2019 was performed, risk factors and pathogens of HAI were analyzed. Results A total of 1 133 patients were monitored, 120 cases (174 times) of HAI occurred, HAI rate was 10.59%, case infection rate was 15.36%; incidence of HAI was 11.95‰ per 1 000 bed-day, case incidence was 17.32‰ per 1 000 bed-day, incidence of HAI adjusted by patient's average severity was 3.65‰ per 1 000 bed-day, case incidence was 5.30‰ per 1 000 bed-day. Utilization rates of respirator, urinary catheter and central venous catheter were 44.25%, 85.19% and 52.19% respectively, incidence of ventilator-associated pneumonia, catheter-associated urinary tract infection and central line-associated bloodstream infection (CLABSI) were 12.37‰, 0.35‰, and 6.10‰ respectively. The main HAI site was lower respiratory tract (64.94%), followed by blood (21.27%) and urinary tract (3.45%). Length of ICU stay ≥ 14 days (OR=0.122, 95% CI: 0.036-0.414, P=0.001), blood transfusion (OR=0.233, 95% CI: 0.104-0.522, P < 0.001), APACHE Ⅱ score ≥ 20 points (OR=0.358, 95% CI: 0.175-0.733, P=0.005) and tracheotomy (OR=0.158, 95% CI: 0.066-0.374, P < 0.001) were independent risk factors for HAI in ICU patients. The mortality of ICU patients with HAI was higher than that of patients without HAI (37.50% vs 28.04%, χ²=4.664, P=0.031). A total of 147 strains of pathogenic bacteria were isolated from 120 patients with HAI, predominantly Gram-negative bacteria (58.50%), the top 5 pathogens of HAI were Klebsiella pneumoniae (19.05%), Candida albicans (17.69%), Pseudomonas aeruginosa (15.65%), Staphylococcus aureus (10.89%) and Acinetobacter baumannii (6.80%). Resistance rates of Klebsiella pneumoniae and Pseudomonas aeruginosa to imipenem were 14.29% and 65.22% respectively, resistance rate of Staphylococcus aureus to oxacillin was 81.25%. Conclusion Incidence of device-associated infection in newly-built general ICU is high, especially CLABSI, antimicrobial resistance of the main pathogens is serious, corresponding prevention and control measures according to the characteristics of related infection should be formulated to reduce the occurrence of HAI.