QUANTITATIVE ASSESSMENT OF HEPATITIS-C VIRUS-RNA BY POLYMERASE CHAIN-REACTION AND A DIGOXIGENIN DETECTION SYSTEM - COMPARISON WITH BRANCHEDDNA ASSAY

A method for quantifying hepatitis C virus (HCV) RNA in serum using re verse transcription-polymerase chain reaction (RT-PCR) followed by slo t-blot hybridization with a specific, digoxigenin-labeled probe was de veloped. Using RNA synthesized from cloned HCV cDNA as a standard, ser um concentration of HCV RNA above 105 copies/ml can be quantitatively determined. To compare this method with branched DNA (bDNA) assay, 45 serum samples from 26 patients with newly acquired acute hepatitis C ( n = 16) or hepatitis C with acute exacerbation (n = 10) were submitted to both assays. HCV RNA in 30 (67%), 12 (27%) and three (6.7%) sample s can be quantitatively determined by both, either and none of the two assays, respectively. Using a standardized qualitative HCV RNA detect ion test (Amplicor HCV test) as a reference, 1 and 0 false positive re sults were found by bDNA and this assay, respectively. This quantitati ve assay using RT-PCR and a digoxigenin detection system was comparabl e to bDNA assay. Since a false positive result was rarely found, this technique can be used as a first line test to screen a large number of samples rapidly and economically. (C) 1997 Elsevier Science B.V.