Detection ofHepatitis C VirusRibonucleic AcidintheSerum byAmplification withPolymerase ChainReaction

Hepatitis C virus (HCV)RNA wasdetected intheseraof patients with non-A, non-B chronic liver disease bypolymerase chain reaction (PCR). RNA wasextracted fromtheserum, reverse transcribed tocDNA,andamplified byPCR.Withthis method, 30patients withnon-A, non-Bchronic liver disease and10healthy subjects weretested. HCV RNA wasdetected in13of16(81%)anti-HCV-positive patients andalsoin7of 14(50%) anti-HCV-negative patients, butinnoneof10antiHCV-negative healthy subjects. Specificity ofthis method was confirmed bydirect sequencing ofamplified cDNAsegment. Thenucleotide sequences (37nucleotides) obtained from15 patients showed only68-78%homology compared withthe prototype HCVnucleotide sequence. Inaddition, of15nucleotide sequences, there were12different types. Butthetranslated aminoacid sequences (12amino acids) showed 83-100% homology compared withtheprototype HCV aminoacid sequence. Thesedatasuggest themajority ofanti-HCV-positive patients arecarriers ofHCV.Buttodetect alltheviremic patients, theanti-HCV antibody testing maybeinsufficient. Direct detection ofHCVRNA maybeuseful inthestudy ofvirus replication anditsassociation withvarious liver diseases. (J. Clin. Invest. 1990. 86:1764-1767.) Keywords: anti-HCV antibody - non-A, non-Bchronic hepatitis - direct sequencingcDNA*hepatocellular carcinoma