A single tube two compartment reverse transcription polymerase chain reaction system for ultrasensitive quantitative detection of hepatitis C virus RNA.
European legislation requiring polymerase chain reaction (PCR) screening of certain plasma pools to exclude contamination with hepatitis C virus (HCV) will be introduced shortly. The concentration of HCV-RNA in contaminated pools is likely to be much lower than that typically found 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 approximately 40 genomes/ml, which represents a sensitivity increase of 10-100 fold over that achieved by currently available assays. This approach to quantitative reverse transcription PCR (RT-PCR) may have wide applications.
Whitby
K
K
Department of Virology, University College London Medical School, UK.
Garson
J A
JA
D016174:Hepacivirus / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012367:RNA, Viral / Q000032:analysis*; D012680:Sensitivity and Specificity
D012367:RNA, Viral