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OBJECTIVE.Contaminated environmental surfaces are an important source for transmission of Multidrug-Resistant Organisms(MDROs).Effective cleaning of surfaces within hospital rooms is necessary to prevent transmission.We investigated the effects of sequential enhanced cleaning measures by culturing high-touch surfaces in a general intensive care unit(GICU).METHODS.A prospective controlled,quasi-experimental study was conducted during a 17-month period in the 25-bed GICU.The study comprised a baseline period(period 1)and 4 sequential tiered interventions periods: each patient zone was wiped with one clean microfiber cloth daily(period 2),fluorescent markers and adenosine triphosphate(ATP)assay were used to monitor and feedback the effectiveness of cleaning(period 3),wiping one patient zone with three clean microfiber cloths daily(period 4)and withdrawal of the feedback from fluorescent markers and ATP assay(period 5).Methicillin-resistant staphylococcus aureus(MRSA),extended-spectrum β-lactamase(ESBL)-producing enterobacteriaceae and vancomycin-resistant enterococci(VRE)were used as marker organisms.Cultures of environmental samples from high-touch surfaces were performed.RESULTS.Compared to period 1,the mean percentages of MDROs positive cultures from high-touch surfaces was reduced by 41.0%(prevalence ratio(OR)=0.59 [95%confidence interval(CI),0.46-0.77],P<0.001),70.8%(OR=0.29,95%CI,0.20-0.43,P<0.001),82.6%(OR=0.17,95%CI,0.12-0.26,P<0.001)and 70.8%(OR=0.29,95%CI,0.21-0.39,P<0.0001)in period 2,3,4 and 5 respectively.Relative to one cloth for a patient zone,wiping each patient zone with three cloths resulted in a statistically significant 41.6%(OR=0.56,95%CI,0.34-0.91;P=0.018)and 50.5%(OR=0.58,95%CI,0.42-0.80;P=0.001)decrease in the prevalence of MDROs positive cultures with or without fluorescent markers and ATP assay.CONCLUSIONS.Enhanced cleaning reduced environmental contamination and using three cleaning cloths for one patient zone was more effective in ensuring effective cleaning of high-touch surfaces when compared to one cleaning cloth.