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Background: Hepatitis C virus (HCV) infection is a global health problem and continues to be a major disease burden in the world, associated with serious health challenges including liver cirrhosis, cancer, lymphomas and death. This study was carried out to determine the prevalence of HCV infection among students of the University of Calabar. Methodology: In a cross-sectional study, 200 students were tested for the presence of anti-HCV antibodies using a rapid immunochromatographic (ICT) assay (CTK Biotech, Inc. USA). Seropositive samples were confirmed using reverse transcriptase-polymerase chain reaction (RT-PCR) assay for detection of HCV RNA. Structured questionnaires were used to collect subjects' socio-demographic data and risk factors of infection. Data were analyzed using SPSS version 16.0, with the level of significance set at p<0.05. Results: Of the 200 students screened, the seroprevalence of HCV was 15.0% (n=30) and 9.5% (n=19) was positive for HCV RNA by RT-PCR assay. The prevalence of anti-HCV antibody was significantly higher in females (18.8%, 12/64) than males (13.2%, 18/136) (x²=3.84, p=0.036). Alcohol consumption (OR=4.67, 95% CI=2.04-10.67, p=0.002), skin piercing (OR=32.99, 95% CI=5.95-72.37, p<0.0001), multiple sexual partners (OR=4.03, 95% CI=1.7-9.6, p=0.0018), and history of blood transfusion (OR=8.00, 95% CI=2.97-21.58, p<0.001) were risk factors significantly associated with HCV infection in the study participants. Conclusion: The findings of 15.0% and 9.5% prevalence of HCV infection by anti-HCV antibody and HCV RNA, respectively in this study, showed that there is relatively high prevalence of HCV infection among the students' population in University of Calabar, Nigeria. Hence, routine medical screening of students for HCV infection using rapid ICT and RT-PCR techniques is hereby recommended. [ABSTRACT FROM AUTHOR]