Detection of hepatitis A virus in a factor VIII preparation by antigen capture/PCR.
The antigen capture/PCR (AC/PCR) has been applied in the analysis of various factor VIII preparations, which were suspected to be contaminated with hepatitis A virus (HAV). AC/PCR involves capturing the antigen, i.e. the intact virus particles, by binding to the HAV monoclonal antibody mAb 7e7, reverse transcription of the viral RNA and amplification of the cDNA with HAV-specific primer pairs. The PCR analysis of one factor VIII concentrate yielded an HAV-specific DNA product, which could be confirmed by Southern blot analysis. The HAV strain recovered by AC/PCR from this factor VIII concentrate could be classified into genotype III after solid-phase sequencing of the product and comparison with the consensus sequences for the known HAV genotypes. Analysis of the sera from 3 haemophiliacs treated with this batch has not resulted in a reliable product. However, the results obtained indicate that HAV can be detected in purified factor VIII preparations by AC/PCR.
Normann
A
A
Department of Medical Virology and Epidemiology of Virus Diseases, University of Tübingen, FRG.
Graff
J
J
Flehmig
B
B
D001483:Base Sequence; D004340:Drug Contamination; D005169:Factor VIII; D006507:Hepatovirus / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods
D005169:Factor VIII