Detection of HCV RNA in Serum by Reverse Transcriptase-PCR and Radiolabeled Liquid Hybridization

Hepatitis C virus (HCV) possesses a single-stranded, positive-sense RNA that is 9.4 kb in length. The complete HCV genome has been cloned and sequenced and encodes for a nucleocapsid, an envelope, and five nonstructural proteins (1,2). The 5' untranslated region of the virus is highly conserved among the HCV genotypes that have been identified to date (3-5) and has been selected by most investigators as the site for developing oligonucleotide primers and probes for the polymerase chain reaction (PCR) (6-9). PCR has proven to be a rapid, sensitive, and useful method for the detection of HCV infections (6-16). The assay can detect HCV in HCV antibody-negative individuals suspected of having hepatitis and can discriminate chronic HCV infections from resolved acute infections in patients who are positive for HCV antibody. The procedure can also be used to: 1. diagnose HCV infections in newborns of HCV-infected women; 2. resolve indeterminate serologic results; 3. monitor antiviral therapy, and 4. identify HCV infection in high-risk, seronegative individuals.