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An improved hydrogen peroxide disinfectant yielded less surface contamination than a quaternary ammonium disinfectant.Improved hydrogen peroxide disinfectant yielded surfaces with no growth more often than a quaternary ammonium disinfectant.A composite health care outcome measure was lower with improved hydrogen peroxide disinfectant than with quaternary ammonium disinfectant.Quaternary ammonium–based (Quat) disinfectants are widely used, but they have disadvantages.This was a 12-month prospective cluster controlled crossover trial. On 4 wards, housekeepers performed daily cleaning using a disinfectant containing either 0.5% improved hydrogen peroxide (IHP) or Quat. Each month, 5-8 high-touch surfaces in several patient rooms on each ward were tagged with a fluorescent marker and cultured before and after cleaning. Hand hygiene compliance rates and antimicrobial usage on study wards were obtained from hospital records. Outcomes included aerobic colony counts (ACCs), percent of wiped surfaces yielding no growth after cleaning, and a composite outcome of incidence densities of nosocomial acquisition and infection caused by vancomycin-resistant enterococci, methicillin-resistant Staphylococcus aureus, and Clostridium difficile infection. Statistical analysis was performed using χ2 test, Fisher exact test, Welch test, and logistic regression methods.Mean ACCs per surface after cleaning were significantly lower with IHP (14.0) than with Quat (22.2) (P = .003). The proportion of surfaces yielding no growth after cleaning was significantly greater with IHP (240/500; 48%) than with Quat (182/517; 35.2%) (P < .0001). Composite incidence density of nosocomial colonization or infection with IHP (8.0) was lower than with Quat (10.3) (incidence rate ratio, 0.77; P = .068; 95% confidence interval, 0.579-1.029).Compared with a Quat disinfectant, the IHP disinfectant significantly reduced surface contamination and reduced a composite colonization or infection outcome.