Background: Hepatitis C virus (HCV) infection is the most common cause of acute or chronic hepatitis in patients on hemodialysis (HD). The purpose of this study was to describe the prevalence of positive HCV RNA and investigate injection drug use as an emerging risk factor in patients with chronic renal disease on HD. Methods: This was a multicenter cross-sectional study with 325 patients with chronic renal disease on HD in the period between August 1, 2005 to August 30, 2006, receiving care at four institutions in the city of Porto Alegre, Southern Brazil. Epidemiological data were collected by means of a structured questionnaire. The following laboratory tests were performed: alanine aminotransferase (ALT), anti-hepatitis C virus antibodies (anti-HCV), and qualitative polymerase chain reaction (PCR). Results: Of 325 patients, 68 had positive HCV RNA results. The comparison between patients with positive and negative PCR results revealed signifi cant differences in duration of HD (mean = 71 versus 52.4 months; p = 0.02); previous blood transfusion (92% versus 72%; p < 0.01); injection drug use (13% versus 0.7%; p < 0.01); anti-HCV positivity at start of HD therapy (60% versus 4%; p < 0.01); and mean ALT value (39 versus 26.5; p < 0.01). Logistic regression analysis showed a positive HCV RNA independently associated to being on HD for more than fi ve years [OR: 2.1 (95% CI 1.2 -3.8)]; previous blood transfusion [OR: 3.7 (95% CI 1.4 -9.5)]; and injection drug use [OR: 22.6 (95% CI 4.2 -119.6)]. Conclusion: Injection drug use was an independent risk factor for HCV infection among chronic renal disease patients on HD.