Objective To screen out high risk factors for multidrug-resistant organisms(MDROs) in intensive care unit (ICU) through failure mode and effect analysis (FMEA), and provide reference for prevention and control. Methods FMEA risk assessment method was used to screen out high risk factors for MDRO healthcare-associated infection(HAI) in ICU, improvement measures were put forward and implemented, effect before and after intervention was compared. Results The screened high-risk events were extra bed for excessive patients, weak consciousness of isolation among cleaners, interns and rotary students. Incidence of MDRO HAI was 4.78% (89/1 861) before intervention and 2.08% (34/1 636) after intervention, difference was statistically significant before and after intervention (χ²=18.76,P<0.05). Incidence of carbapenem-resistant Acinetobacter baumannii (CRAB) HAI was 0.31% after intervention, which was lower than 1.40% before intervention (P<0.05); there was no significant difference in HAI rates of methicillin-resistant Staphylococcus aureus, extended-spectrum β-lactamases (ESBLs)-producing Escherichia coli, ESBLs-producing Klebsiella pneumoniae and carbapenem-resistant Enterobacteriaceae before and after intervention (all P>0.05). Conclusion FFMEA can guide department of HAI management and ICU to find out the weak links in the prevention and control of MDRO infection, and carry out targeted prevention and control measures.