title: Applications of quantitative PCR in the biosafety and genetic stability assessment of biotechnology products.
abstract: High throughput screening, increased accuracy and the coupling of real-time quantitative PCR (Q-PCR) to robotic set-up systems are beginning to revolutionise biotechnology. Applications of Q-PCR within biotechnology are discussed with particular emphasis on the following areas of biosafety and genetic stability testing: (a) determination of the biodistribution of gene therapy vectors in animals; (b) quantification of the residual DNA in final product therapeutics; (c) detection of viral and bacterial nucleic acid in contaminated cell banks and final products; (d) quantification of the level of virus removal in process validation viral clearance studies; (e) specific detection of retroviral RT activity in vaccines with high sensitivity; and (f) transgene copy number determination for monitoring genetic stability during production. Methods employed for Q-PCR assay validation as required in ICH Topic Q2A Validation of Analytical Methods: Definitions and Terminology (1st June 1995) are also reviewed.
authors: Q-One Biotech Ltd., West of Scotland Science Park, Glasgow, Scotland, UK. alovatt@q-one.co.uk
authors: Archie
authors: A
authors: Lovatt
mesh_terms: D001709:Biotechnology / Q000379:methods*; D004247:DNA / Q000032:analysis*; D017931:DNA Primers; D004279:DNA, Viral / Q000032:analysis; D004259:DNA-Directed DNA Polymerase / Q000032:analysis; D005092:Exonucleases / Q000032:analysis; D005456:Fluorescent Dyes / Q000378:metabolism; D018628:Gene Dosage; D010957:Plasmids / Q000032:analysis; D016133:Polymerase Chain Reaction; D016415:Sequence Alignment; D019076:Transgenes; D014780:Viruses / Q000378:metabolism
chemical_list: D017931:DNA Primers; D004279:DNA, Viral; D005456:Fluorescent Dyes; D004247:DNA; D004259:DNA-Directed DNA Polymerase; D005092:Exonucleases