Detection of hepatitis C virus in paraffin-embedded liver biopsies of patients negative for viral RNA in serum

Hepatology. 1999 Jan;29(1):223-9. doi: 10.1002/hep.510290118.


The diagnosis of hepatitis C is based on serological testing for antibodies against various epitopes of the hepatitis C virus (HCV) and detection of HCV RNA in serum, because anti-HCV antibodies alone cannot discriminate patients who are infectious from those who have resolved the infection. If HCV RNA is not detected, which is the case in at least 20% of enzyme immunoassay (EIA)-positive patients, diagnosis remains unclear in a state of disease possibly well suited for therapeutic intervention. Therefore, we investigated if detection of HCV antigens or HCV RNA in routinely processed, formalin-fixed and paraffin-embedded (ffpe) liver biopsy specimens of patients positive for anti-HCV, but negative for HCV RNA in serum, could confirm diagnosis in this serological constellation. We detected HCV RNA by reverse-transcription polymerase chain reaction (RT-PCR) in 27 (61%) of 44 ffpe liver biopsies from EIA-positive, but HCV-RNA-seronegative, patients. Testing of 18 of these biopsies by a panel of polyclonal antibodies against structural and nonstructural HCV proteins revealed positive immunostaining in 6 cases (33%), which were also positive by RT-PCR. Most biopsies showed necroinflammation compatible with chronic hepatitis C, and the detection of tissue HCV RNA correlated significantly with a higher grade of inflammatory activity. Detectability of HCV RNA did not correlate with HCV subtype. In conclusion, the search for HCV RNA by RT-PCR within the liver biopsy specimen can establish rapid and unequivocal diagnosis of hepatitis C in at least 60% of anti-HCV antibody-positive patients who are seronegative for HCV RNA, and thus may help to avoid repeated testing and delayed therapy. Tissue RT-PCR may also be more efficient than serological testing for surveillance of interferon therapy response, because ongoing chronic active hepatitis C is clearly demonstrated in the absence of detectable serum HCV RNA.

Publication types

  • Clinical Trial

MeSH terms

  • DNA Primers
  • Hepacivirus / chemistry*
  • Hepacivirus / isolation & purification
  • Hepatitis C Antigens / analysis
  • Humans
  • Immunohistochemistry
  • Liver / immunology
  • Liver / metabolism
  • Liver / virology*
  • Paraffin Embedding
  • RNA, Messenger / biosynthesis
  • RNA, Viral / biosynthesis*
  • RNA, Viral / blood
  • RNA, Viral / genetics
  • Retrospective Studies
  • Reverse Transcriptase Polymerase Chain Reaction
  • Viral Proteins / biosynthesis


  • DNA Primers
  • Hepatitis C Antigens
  • RNA, Messenger
  • RNA, Viral
  • Viral Proteins