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

Abstract

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.

• Authors: V Dries, I von Both, M Müller, G Gerken, P Schirmacher, M Odenthal, R Bartenschlager, U Drebber, K H Meyer zum Büschenfeld, H P Dienes
• Journal: Hepatology (Baltimore, Md.) (Hepatology) Vol. 29 Issue 1 Pg. 223-9 (Jan 1999) ISSN: 0270-9139 [Print] United States
• PMID: 9862870 (Publication Type: Clinical Trial, Journal Article)

Chemical References

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

Topics

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