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Detection of hepatitis "C" virus in formalin-fixed liver tissue by nested polymerase chain reaction.

作者信息

Sallie R, Rayner A, Portmann B, Eddleston A L, Williams R

机构信息

Institute of Liver Studies, King's College School of Medicine and Dentistry, King's College Hospital, London, England.

出版信息

J Med Virol. 1992 Aug;37(4):310-4. doi: 10.1002/jmv.1890370415.

Abstract

Interpretation of antibody to hepatitis C virus (HCV) in patients with liver disease is difficult due to false-positive reactivity in some conditions. To evaluate the feasibility of HCV in archival material, HCV was sought in formalin-fixed, paraffin-embedded liver biopsy specimens. Nested polymerase chain reaction was used to detect hepatitis C virus in formalin-fixed, paraffin-embedded liver biopsy specimens after total RNA was extracted from tissue by proteinase K digestion and phenol/chloroform purification. The relative efficiency of amplification of HCV RNA from formalin-fixed material was estimated semiquantitatively by serial dilution of cDNA synthesised from RNA extracted from fresh and formalin-fixed sections from the same liver. Although HCV RNA could be detected in formalin-fixed liver tissue by nested PCR in 5/5 cases in which HCV was detected in serum, amplification was approximately 5-fold less efficient than when HCV was amplified from fresh tissue. Nevertheless, nested PCR of HCV from formalin-fixed liver tissue represents a useful technique in addressing some important questions related to the pathogenesis of liver disease.

摘要

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