Background: Hepatitis C virus (HCV) infection remains a significant public health problem worldwide, with varying epidemiological trends over time necessitating continual surveillance and intervention strategies. Objectives: This retrospective study aimed to examine trends in demographics, transmission routes, and disease characteristics among 635 patients diagnosed with HCV infection between 2003 and 2023. Methods: Data on demographics, HCV RNA levels, liver enzymes, genotype distribution, and transmission routes were retrospectively analyzed. The study period was divided into four segments for detailed analysis. Results: There was a gradual increase in the diagnosed cases over the study period, particularly noticeable after 2012. The mean age at diagnosis remained stable from 2003 - 2007 to 2008 - 2012 but decreased significantly in the following years (P = 0.001). Male predominance was observed throughout the study, with a notable increase in the male/female ratio over time (P < 0.001). Intravenous drug use (IVDU) emerged as the predominant transmission route, reaching 68.3% in 2018 - 2022 (P < 0.001). The prevalence of incarceration history among patients increased significantly, reaching 24.6% in 2018 - 2022 (P < 0.001). Disease characteristics varied between periods; median HCV RNA levels were highest in 2018 - 2022, and median alanine aminotransferase (ALT) levels differed significantly between periods (P = 0.001). The genotype distribution showed a shift over time, with genotype 1 predominating initially and genotypes 3, 4, and 5 becoming more prevalent in later years (P < 0.001). Conclusions: This study highlights the dynamic changes in hepatitis C epidemiology over two decades in southern Turkey, highlighting the increasing burden in younger populations, the increasing prevalence of IVDU as a transmission route, and the changing genotype distribution.