Covalently closed circular DNA (cccDNA) is a unique episomal replicative intermediate molecule of hepatitis B virus (HBV) which plays a key role in viral persistence. The aim of this study was to prove cccDNA persistence in the liver tissue of patients negative for hepatitis B surface antigen (HBsAg) and positive for antibody to hepatitis core antigen (anti-HBc). Intrahepatic HBV DNA and cccDNA were determined using real-time and semi-nested PCR assays on the liver tissues of 35 patients who were negative for HBsAg and positive for anti-HBc with or without anti-HBs. HBV DNA was detected in the liver tissue of 4 out of 35 patients who were positive for anti-HBc. None of the samples harbored cccDNA. In this study population, which is of Asian origin, very low levels of HBV DNA were detected in a small percentage of patients with anti-HBc. Even using the highly sensitive semi-nested PCR assay, HBV cccDNA was not detectable in any anti-HBc positive patients either with or without anti-HBs.