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BACKGROUND AND OBJECTIVES: Screening for hepatitis C virus (HCV) by polymerase chain reaction (PCR) will be mandatory for screening blood and plasma donors in Europe and elsewhere. This study describes an internally controlled, highly sensitive PCR method designed for screening blood donations in pools. MATERIAL AND METHODS: RNA extracted from bovine viral diarrhoea virus (BVDV) was used as an internal control to monitor the efficiency of extraction, reverse transcription and amplification steps in HCV PCR. RESULTS: Sensitivity of PCR for single molecules of HCV in the presence of 33 genome equivalents of BVDV RNA was achieved by reducing the efficiency of BVDV amplification. BVDV could be recovered at high efficiency from large volume pools (2-5 ml) by ultracentrifugation and by the NucliSens extraction method. CONCLUSION: Detection of BVDV validates the extraction, reverse transcription and amplification methods used for HCV detection in plasma pools and provides valuable quality assurance for negative results.

Original publication

DOI

10.1159/000031044

Type

Journal article

Journal

Vox Sang

Publication Date

1999

Volume

76

Pages

170 - 174

Keywords

Animals, Blood Donors, Cattle, Chemical Phenomena, Chemistry, Physical, Diarrhea Viruses, Bovine Viral, Hepacivirus, Mass Screening, Reference Standards, Reverse Transcriptase Polymerase Chain Reaction, Sensitivity and Specificity