Serum samples with lower temperature-dependent antibody with low avidity to C100-3 (HCV) antigen were found in 0.19% of 23,197 voluntary blood donors at this blood center. They showed positive C100-3 antibody activity at 24 C but not at 37 C. The antibody activity bound to C100-3 antigen at lower temperature disappeared after incubation for 60 min at 37 C or treatment with 8 M urea. Other markers of hepatitis C virus infection, especially the presence of HCV-RNA were demonstrated in some of these serum samples and the importance of this phenomenon is discussed with regard to virus screening of blood donors for hepatitis C.