PCR ANALYSIS OF HEPATITIS-B VIRUS-DNA IN PARAFFIN-EMBEDDED LIVER-TISSUE FROM PATIENTS WITH CHRONIC LIVER-DISEASE

We described a nested polymerase chain reaction protocol to detect hep atitis B viral DNA in paraffin-embedded liver tissue and tried to dete rmine whether this virus was associated with non-B chronic liver disea se. Fifty-five samples were obtained from 28 patients with B, and 27 p atients with non-B chronic liver disease (35 cirrhosis, 4 hepatocellul ar carcinoma and 16 chronic hepatitis). The two sets of primers amplif y a sequence located in a conserved polymerase/surface region of the v iral genome. Reaction products were analysed using a nonisotopic hybri dization method. None of the 27 (0%) seronegative samples and 20 of th e 28 (71%) seropositive specimens were positive for hepatitis B virus DNA. There were 4 false negatives in which beta-globin PCR was positiv e. Although its sensitivity is reduced in formalin-fixed paraffin-embe dded tissue, nested PCR allows rapid detection of HBV DNA sequences an d can be a useful tool if no frozen tissue is available.