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Detection of Hepatitis A Virus in a Factor VIII Preparation by Antigen Capture/PCR
Author(s) -
Normann Andrea,
Graff Judith,
Flehmig Bertram
Publication year - 1994
Publication title -
vox sanguinis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.68
H-Index - 83
eISSN - 1423-0410
pISSN - 0042-9007
DOI - 10.1111/j.1423-0410.1994.tb00979.x
Subject(s) - virology , antigen , biology , monoclonal antibody , microbiology and biotechnology , virus , genotype , complementary dna , primer (cosmetics) , antibody , reverse transcriptase , polymerase chain reaction , chemistry , gene , immunology , genetics , organic chemistry
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.