This study aimed to analyze temporal trends by gender and age in Clostridium difficile infection (CDI)-related hospitalization rates in the Autonomous Community of Madrid (Spain) over a 12-year period. A population-based cross-sectional study of all hospital admissions with a CDI diagnosis from 2003 to 2014 was carried out. Annual age-specific hospitalization rates were calculated by gender. All the analyses were performed separately for total hospitalizations and hospitalizations with CDI as the primary diagnosis. Joinpoint regression models were used to analyze time trends. A total of 13,526 hospital discharges were identified (26.8 % with CDI as the primary diagnosis). In both sexes, a gradient in age-specific rates was observed, ranging in 2014 from 5.92 hospitalizations per 100,000 person-years in patients <15 years of age to 378.96 in patients ≥85 years of age. Since 2009, in the age group of 15-44 years, both men and women presented an increasing trend of around 18 %. A significantly increasing trend was detected in women of age 45-84 years, with an estimated annual percentage of change of 7.6 % in the age group of 45-64 years, and rounding with 4.5 % in the age group of 65-84 years. In men of age 45-64 years, the average annual percentage of increase was 4.7 %, and it was 21.1 % between 2010 and 2014 in the age group of 65-74 years. No trends were identified in the 85 years and over age group. Surveillance methods to assess trends by age group should be implemented. Preventive and therapeutic initiatives should remain a priority.