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 hepatitis B viral DNA in paraffin-embedded liver tissue and tried to determine whether this virus was associated with non-B chronic liver disease. Fifty-five samples were obtained from 28 patients with B, and 27 patients with non-B chronic liver disease (35 cirrhosis, 4 hepatocellular carcinoma and 16 chronic hepatitis). The two sets of primers amplify a sequence located in a conserved polymerase/surface region of the viral genome. Reaction products were analysed using a nonisotopic hybridization method. None of the 27 (0%) seronegative samples and 20 of the 28 (71%) seropositive specimens were positive for hepatitis B virus DNA. There were 4 false negatives in which beta-globin PCR was positive. Although its sensitivity is reduced in formalin-fixed paraffin-embedded tissue, nested PCR allows rapid detection of HBV DNA sequences and can be a useful tool if no frozen tissue is available.
Chapel
F
F
Department of Pathology, Hôpital de la Timone, Marseille, France.
de Lamballerie
X
X
de Micco
C
C
Lebreuil
G
G
de Micco
P
P
D000293:Adolescent; D000328:Adult; D000368:Aged; D001483:Base Sequence; D006528:Carcinoma, Hepatocellular / Q000821:virology; D002908:Chronic Disease; D004279:DNA, Viral / Q000302:isolation & purification*; D004587:Electrophoresis, Agar Gel; D005188:False Negative Reactions; D005260:Female; D006515:Hepatitis B virus / Q000235:genetics*; D006801:Humans; D008099:Liver / Q000821:virology; D008107:Liver Diseases / Q000821:virology*; D008113:Liver Neoplasms / Q000821:virology; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D016612:Paraffin Embedding; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D012699:Serology
D004279:DNA, Viral