We investigated hepatitis B virus (HBV) DNA status in the liver of 22 patients with hepatocellular carcinoma (HCC) developed on a non-cirrhotic, histologically normal liver tissue. HBV serological markers were present in 2 of the 22 subjects. HBV DNA sequences were identified in the liver of only 5 of the 22 patients with HCC. Evidence for clonal expansion of HBV-infected cells was found for one HBsAg-positive subject. This study indicates a much lower rate of HBV DNA positivity in the group of HCC developed on histologically normal livers as compared to that observed in HCC with liver cirrhosis.