A SINGLE TUBE 2 COMPARTMENT REVERSE-TRANSCRIPTION POLYMERASE-CHAIN-REACTION SYSTEM FOR ULTRASENSITIVE QUANTITATIVE DETECTION OF HEPATITIS-C-VIRUS-RNA

European legislation requiring polymerase chain reaction (PCR) screeni ng of certain plasma pools to exclude contamination with hepatitis C v irus (HCV) will be introduced shortly. The concentration of HCV-RNA in contaminated pools is likely to be much lower than that typically fou nd in single donations, so the sensitivity of the PCR assays used for the screening will be critical. We describe here the development of a quantitative PCR assay for HCV-RNA with a detection limit of approxima tely 40 genomes/ml, which represents a sensitivity increase of 10-100 fold over that achieved by currently available assays, This approach t o quantitative reverse transcription PCR (RT-PCR) may have wide applic ations. (C) 1997 Elsevier Science B.V.