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• W2075749649 startingPage "476" @default.
• W2075749649 abstract "The genetic polymorphism of the cytochrome P450 monooxygenase, CYP2D6, comprises at least 43 alleles giving rise to distinct drug metabolism phenotypes termed ultrarapid, extensive, intermediate, and poor metabolizers. As a consequence, drug side effects or lack of drug effect may occur if standard doses are applied. Genetic prediction of drug oxidation phenotype as a basis for dose selection requires analysis of single nucleotide polymorphisms and of alleles with duplicated or deleted genes. Here we developed a novel method to determine the CYP2D6 gene dose per genome. A TaqMan real-time PCR assay to specifically amplify genomic CYP2D6 was established by using a specific set of amplification primers and probe, located in exon 9, which effectively prevent amplification of CYP2D7 and CYP2D8 pseudogenes. Quantitative CYP2D6 amplification data were normalized to albumin as an internal reference gene which was coamplified simultaneously in a single-tube biplex assay. The assay was validated with a selection of previously genotyped DNA samples containing none, one, two, or three CYP2D6 gene copies. The results were highly reproducible and closely matched the number of genes with no overlap between the groups. Analysis of DNA samples comprising all major alleles and genotypes revealed high sensitivity and specificity of the assay, as demonstrated by agreement of the determined gene dose with the presence of CYP2D6(*)2 x 2 (gene duplication) and CYP2D6(*) 5 (gene deletion) alleles. The predictability of the new strategy was systematically evaluated. The semiautomatic TaqMan assay allows high sample throughput and will be useful for pharmacogenetic studies and in the clinical setting." @default.
• W2075749649 created "2016-06-24" @default.
• W2075749649 creator A5023657124 @default.
• W2075749649 creator A5044688765 @default.
• W2075749649 creator A5057183308 @default.
• W2075749649 creator A5066057635 @default.
• W2075749649 date "2003-11-17" @default.
• W2075749649 modified "2023-10-01" @default.
• W2075749649 title "CYP2D6 genotyping strategy based on gene copy number determination by TaqMan real-rime PCR" @default.
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• W2075749649 doi "https://doi.org/10.1002/humu.10280" @default.
• W2075749649 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/14635107" @default.
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