Transfusion-transmitted virus (TTV) has been identified from patients with post-transfusion hepatitis of unknown aetiology, but the clinical relevance remains unclear. The aim of this study was to evaluate TTV in liver. We studied 15 patients with hepatitis non-A-E and 44 with hepatitis B virus (HBV). The nested polymerase chain reaction (PCR) with primers corresponding to the conserved region of the published TTV genome was employed to amplify TTV fragments in serum, and in situ hybridization was used to detect TTV in biopsied liver specimens. TTV DNA was detected in serum from six (40%) of 15 patients with hepatitis of unknown aetiology and from 16 (36.4%) of 44 patients with chronic hepatitis B, respectively. The intrahepatic viral fragment was detected in 17 (77.3%) of 22 patients with TTV in serum. There was no statistical difference in the prevalence of TTV infection between the two groups (hepatitis non-A-E 40% vs HBV 25%, P > 0.75). When patients in both groups, with and without TTV, were compared, no differences were found in serum alanine aminotransferase (ALT) levels (hepatitis non-A-E: 131.5 +/- 66.6 vs 244.2 +/- 257.4, P=0.955; HBV: 240.1 +/- 418.9 vs 214.6 +/- 276.7 U l(-1), P=0.761) or histological activity index (grade) score (hepatitis non-A-E: 6.4 +/- 5.5 vs 5.6 +/- 5.9, P=0.689; HBV: 5.6 +/- 3.7 vs 5.5 +/- 3.7, P=0.345). HBV DNA levels in patients with and without TTV co-infection did not differ significantly (300 +/- 776.4 microg ml(-1) vs 97.1 +/- 160.5 microg ml(-1), P=0.980). Hence, TTV does exist in liver, but plays no role in hepatitis or aggravation of liver damage when co-infected with HBV.