Five hundred serum samples from volunteers blood donors were investigated in order to determine the presence of anti-HCV antibodies by an enzyme-linked immunoassay recently developed worldwide. Prevalence of true-positive samples was 1.2% (6/500), 2 fold higher than the reported prevalence in most of the industrialized countries. From these 6 sera, one (16.6%) showed simultaneous reactivity for HBV anticore antibody. Three sera (25%) from 12 patients with diagnosis of Non-A Non-B hepatitis were reactive for anti-HCV antibodies while in a group of 32 sera with negative HAV and HBV screening, 4 (12.5%) showed anti-HCV antibodies. Two samples out of 16 sera persistently positive for anticore demonstrated the presence of anti-HCV antibodies. The anti-HCV antibodies were undetectable in two cases of autoimmune chronic hepatitis. Our results indicate that in Venezuela, HCV represents a significant problem of public health coexisting in certain cases with HBV infection.