The aim of this cross-sectional seroprevalence study was to determine the prevalence of antibodies to hepatitis C virus (HCV) (anti-HCV) in patients with cirrhosis, hepatocellular carcinoma (HCC) and chronic active hepatitis (CAH) attending a referral hospital in a hepatitis B virus (HBV)-endemic area in South Africa. One hundred and ten patients with suspected cirrhosis, 44 with suspected HCC and 6 with chronic hepatitis were initially included. The diagnoses were confirmed in 77 patients with cirrhosis (histologically or macroscopically at peritoneoscopy), 33 patients with HCC (histologically or elevated alpha-fetoprotein levels plus focal lesion on hepatic imaging) and 6 patients with CAH (histologically) without antinuclear antibodies. All patients were tested for anti-HCV with the Abbott second-generation enzyme immunoassay combined with a supplemental neutralisation assay, and hepatitis B surface antigen (HBsAg). Anti-HCV seroprevalence for cirrhosis, HCC and CAH were 18/77 (23%), 8/33 (24%) and 2/6 (33%) respectively. HBsAg was detected in serum in 16 (21%), 15 (46%) and 1 (17%) patient respectively. Only 1 patient (with cirrhosis) was positive for both anti-HCV and HBsAg. Of those who were anti-HCV-positive, 4/18 (22.2%) cirrhotics, none with HCC and 1/2 (50%) with CAH, had previously received blood transfusions, resulting in a cumulative frequency of 5/28 (18%). Our results indicate that HCV is an important aetiological agent in the pathogenesis of chronic liver disease in our patients. In the majority of patients (82%), the infection was not transfusion-related. Thus, screening of blood donors for anti-HCV would not prevent the majority of cases of chronic liver disease secondary to HCV. It appears as if HCV and HBV have different modes of transmission in southern Africa.