Use of bovine viral diarrhoea virus as an internal control for amplification of hepatitis C virus.
OBJECTIVE
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.


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.


CONCLUSIONS
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.
Cleland
A
A
Scottish National Blood Transfusion Service Microbiology Reference Laboratory, International Research Centre, Penicuik, UK.
Nettleton
P
P
Jarvis
L
L
Simmonds
P
P
D000818:Animals; D001782:Blood Donors; D002417:Cattle; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D001908:Diarrhea Viruses, Bovine Viral / Q000235:genetics*; D016174:Hepacivirus / Q000235:genetics*; D008403:Mass Screening / Q000379:methods*; D012015:Reference Standards; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity