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Detection of hepatitis “C” virus in formalin‐fixed liver tissue by nested polymerase chain reaction
Author(s) -
Sallie Richard,
Rayner Anne,
Portmann Bernard,
Eddleston A. L. W. F.,
Williams Roger
Publication year - 1992
Publication title -
journal of medical virology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.782
H-Index - 121
eISSN - 1096-9071
pISSN - 0146-6615
DOI - 10.1002/jmv.1890370415
Subject(s) - nested polymerase chain reaction , hepatitis c virus , virology , polymerase chain reaction , virus , liver disease , biology , hepatitis , liver biopsy , rna , antibody , hepatitis c , proteinase k , pathology , biopsy , microbiology and biotechnology , medicine , dna , immunology , gene , biochemistry , genetics
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.