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BACKGROUND The development of hospital-onset Clostridioides difficile infection (HO-CDI) is affected by patient and environmental risk factors. We investigated changes in the incidence of HO-CDI after relocation to a newly built hospital with 50% private rooms and evaluated the associated factors. METHODS A retrospective study was conducted to assess trends in CDI incidences before and after the relocation using segmented regression analysis model. The association between CDI incidence and environmental factors at the ward-level was assessed using a linear regression analyses model. RESULTS The HO-CDI incidence decreased from 6.14 to 1.17 per 10,000 patient-days in the old and new hospital, respectively. Similarly, the community-onset CDI (CO-CDI) incidence decreased from 1.71 to 0.46 per 1000 admissions. HO-CDI incidence was positively correlated with CO-CDI incidence and inversely correlated with the private room ratio (adjusted R2=0.83). Almost half of the CO-CDI patients had been hospitalized within 28 days preceding the onset. DISCUSSION Environmental improvements after relocation may have reduced the reservoir of C. difficile, resulting in a decrease in the number of asymptomatic carriers and CO-CDI patients. CONCLUSION Relocation to a new hospital significantly reduced HO-CDI incidence, concomitantly decreasing the incidence of CO-CDI, potentially due to environmental improvements.