ABSTRACT: OBJECTIVE:Clostridioides difficile infection (CDI) is rapidly increasing in children's hospitals nationwide. Thus, we aimed to compare the effectiveness of 9 infection prevention interventions and 6 multiple-intervention bundles at reducing hospital-onset CDI and asymptomatic C. difficile colonization. DESIGN:Agent-based simulation model of C. difficile transmission. SETTING:Computer-simulated, 80-bed freestanding, tertiary-care pediatric hospital, including 8 identical wards with 10 single-bed patient rooms each. PARTICIPANTS:The model includes 5 distinct agent types: patients, visitors, caregivers, nurses, and physicians. INTERVENTIONS:Daily and terminal environmental disinfection, screening at admission, reduced intrahospital patient transfers, healthcare worker (HCW), visitor, and patient hand hygiene, and HCW and visitor contact precautions. RESULTS:The model predicted that daily environmental disinfection with sporicidal product, combined with screening for asymptomatic C. difficile at admission, was the most effective 2-pronged infection prevention bundle, reducing hospital-onset CDI by 62.0% and asymptomatic colonization by 88.4%. Single-intervention strategies, including daily disinfection, terminal disinfection, asymptomatic screening at admission, HCW hand hygiene, and patient hand hygiene, as well as decreasing intrahospital patient transfers, all also reduced both hospital-onset CDI and asymptomatic colonization in the model. Visitor hand hygiene and visitor and HCW contact precautions were not effective at reducing either measure. CONCLUSIONS:Hospitals can achieve substantial reduction in hospital-onset CDIs by implementing a small number of highly effective interventions.