Objective To evaluate the effect of environmental screening of multidrug-resistant organisms (MDROs) on reducing methicillin-resistant Staphylococcus aureus (MRSA) and carbapenem-resistant Acinetobacter baumannii (CRAB) on object surface in intensive care unit (ICU). Methods Baseline survey period was from September to November in 2018, intervention period was from December 2018 to August 2019, during intervention period, routine screening was conducted every quarter, timely feedback was given, environmental cleaning and disinfection as well as related knowledge training was strengthened; isolation of MRSA and CRAB on surface of environmental objects before and after intervention were compared. Results 896 specimens were collected during baseline survey period, 2 880 specimens were collected after intervention, MRSA isolation rate decreased from 18.08% to 0, CRAB isolation rate decreased from 4.46% to 0.83%; isolation rates of MRSA and CRAB in hospitalized patients were 48.86% and 36.08% respectively at baseline survey period, isolation rates of MRSA and CRAB were 37.42% and 27.27% respectively after intervention; isolation rates of MRSA and CRAB isolated from environmental object surface and hospitalized patients before and after intervention were both significantly different (both P < 0.05), there was a trend of gradual decline every quarter. The most common isolation sites of MRSA and CRAB from environment were: bedside floor (34/168, 20.24%), head pad (19/168, 11.31%), ventilator panel (13/168, 7.74%), bedside table (12/168, 7.14%), and medical hanging tower (12/168, 7.14%). At baseline survey period, isolation rates of MRSA and CRAB in respiratory ICU were the highest, which were 75.00% (24/32) and 25.00% (8/32) respectively, after intervention, MRSA was not isolated in respiratory ICU, isolation rate of CRAB was 3.13% (3/96). Conclusion Routine environmental screening for MDROs in key departments can effectively reduce the colonization of MDROs on the surface of ICU objects and reduce the risk of infection in patients.