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

✍ Scribed by Richard Sallie; Anne Rayner; Bernard Portmann; A. L. W. F. Eddleston; Dr. Roger Williams

Publisher
John Wiley and Sons
Year
1992
Tongue
English
Weight
519 KB
Volume
37
Category
Article
ISSN
0146-6615

No coin nor oath required. For personal study only.

✦ Synopsis

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 ∼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. © 1992 Wiley‐Liss, Inc.

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