Clostridium difficile infection (CDI) is a common and serious nosocomial infection worldwide. It is also problem in our hospital, a medical school in Khon Kaen University. Rationale use of antimicrobial in the treatment of CDI is still being questionable. This study aimed to explore the characteristic and evaluate the treatment of CDI in our setting. This retrospective cohort study was performed in 168 CDI patients admitted at Srinagarind Hospital between October 2013 and September 2016. Patient medical records were reviewed for demographics, comorbidities, antimicrobial exposures, treatments and clinical outcomes. Antimicrobial therapy for CDI was then evaluated in accordance with the current CDI guideline. Most CDI patients aged over 50 years (72.6%) and 42.8% aged over 65 years. Common comorbidities in the patients were cardiovascular disease (47.6%), diabetes mellitus (27.4%), chronic kidney disease (20.8%), malignancy (16.7%), cerebrovascular disease (11.3%), chronic hepatitis (3.6%) and chronic obstructive pulmonary disease (2.4%). Patients who had taken antimicrobial during hospitalization prior to CDI diagnosis constituted 83.9% of total study population. Metronidazole, vancomycin and both were initial therapy in 83.9, 4.8 and 11.3% of CDI, respectively. Appropriateness of CDI treatment in all terms was 44% and recovery rate was 62% of assessable cases. Nine patients (5.4%) died during hospitalization. In conclusion, most patients had CDI risk factors including elderly, comorbidity, antimicrobial exposure, and prolonged hospital stay. Inappropriate CDI treatment was found in asymptomatic carriers or inappropriate treatment duration. Initial therapy with metronidazole should change to oral vancomycin which was the first-line therapy in the current guidelines.