Sera of 40 patients with posttransfusion non-A, non-B hepatitis were tested for hepatitis C and B viral genomes by polymerase chain reaction and for hepatitis C antibodies by synthetic peptide immunoassays. Five were then considered to have chronic hepatitis before transfusion. Six patients without hepatitis C markers and hepatitis B virus DNA recovered. In 29 recipients who became positive for hepatitis C virus RNA, posttransfusion hepatitis C was diagnosed. Of them, 5 were hepatitis B surface antigen carriers. Synthetic peptide immunoassays detected 28 whereas anti-C100 assay detected 23 of the 29 acute hepatitis C patients. Anticapsid antibody appeared earlier than the antinonstructural antibody in 10 seroconverters. They appeared simultaneously in 15 seroconverters but anticapsid antibody appeared later then the antinonstructural antibody in 3 hepatitis B carriers. Transient suppression of hepatitis B surface antigenemia was found in 2, whereas elevated hepatitis B virus DNA was found in 3 carriers during acute hepatitis C superinfection. In 2 carriers whose hepatitis C became chronic, both hepatitis B and C viral genomes persisted throughout 2 years of followup. Therefore these assays define posttransfusion hepatitis C more precisely, and there seems no significant interference between chronic hepatitis B and C virus infections.