Background: Clostridioides difficile infection (CDI), one of the most common healthcare-associated (HCA) infections, primarily affect the elderly and immune-compromised patients following antimicrobial consumption. Methods and Objectives: The aim was to determine the HCA CDI incidence trend, characteristics of patients, and classes of previously used antibiotics in the Institute for Pulmonary Diseases of Vojvodina (IPBV) from 2015 to 2023. The incidence of HCA CDI was calculated per 1,000 hospital admissions. Retrospective analysis of demographic and clinical characteristics of HCA CDI patients was performed by SPSS v24.0. computer software program. Statistical significance was set at p<0.05. Results: During the observed period the total number of hospital admissions was 59,943, and the total number of patients with HCA CDI was 364. The incidence of HCA CDI increased from 3.1/1000 in 2019 to 8.5/1000 in 2023. HCA CDI was significantly more common among males compared to females (60.4% vs. 39.4%, p<0.001). The average age of CDI patients was 68 years (67.87±12.62), with no differences by sex (p=0.241). Antibiotics prior development of CDI were reported in 94.2% of CDI patients, among 76 had 4 to 7 classes of antibiotics. Frequencies of 3th and 4th generation of cephalosporins, fluoroquinolones, macrolides, carbapenems, and beta-lactam antimicrobials were respectively 66.8%, 51.4%, 22.8%, 19.5%, and 16.8%. Prior use of 4 or more classes of antibiotics was significantly common among males compared to females (73.7% vs. 26.3%, p=0.027). The average duration of hospitalization prior to the onset of CDI was 15 days (14.66±10.24), without significant differences by age, gender, number of antibiotics, year of observation, Clinic type or patient outcome. Conclusion: The CDI incidence increased from 2015 to 2023. Most CDI patients were elderly males with more often prior use of 4 or more classes of antibiotics. Hospitalization longer than 15 days was an important risk factor for our CDI patients.