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Objective: To conduct an exploratory study to evaluate the clinical effectiveness and cost benefit of universal versus targeted screening for methicillin-resistant Staphylococcus aureus (MRSA) to prevent hospital-acquired MRSA infections., Design: Prospective, interventional study, using a case-control design, difference-in-differences, and cost-benefit analyses., Setting: Two community hospitals in Wisconsin., Patients: Consecutive sample of 15,049 adult admissions from April 2009 to July 2010., Interventions: MRSA surveillance performed by polymerase chain reaction (PCR) on samples collected from all adult patients (aged over 18 years) within 30 days before or upon an admission to the hospital. During a 9-month baseline period, targeted screening was conducted at both hospitals. During the 5-month intervention period, all patients admitted to the intervention hospital were screened for MRSA. Infection control measures were consistent at both hospitals., Results: Universal screening was associated with an increase in admission screening of 43.58 percentage points (P< .01), an increase in MRSA detection of 2.95 percentage points (P< .01), and a small, nonsignificant decline in hospital-acquired MRSA infections of 0.12 percentage points (P< .01). The benefit-to-cost ratio was 0.50, indicating that for every dollar spent on universal versus targeted screening, only $0.50 is recovered in avoided costs of hospital-acquired MRSA infection., Conclusion: Compared with targeted screening, universal screening increased the rate of detection of MRSA upon hospital admission but did not significantly reduce the rate of hospital-acquired MRSA infection. Universal screening was associated with higher costs of care and was not cost beneficial.