title: Characterization of cytochrome P450 2D6 alleles using the Invader system.
abstract: The cytochrome p450 (CYP) superfamily comprises enzymes that play an essential role in the transformation of medically relevant compounds. Accurate genotyping of polymorphisms in members of this family is drawing increasing interest because certain allelic variants may result in either loss of efficacy or toxic accumulation of therapeutic agents. Debrisoquine 4-hydroxylase, or CYP2D6, is among the most widely studied of the CYPs. The complexity of the CYP2D6 genomic region, including pseudogenes, gene deletions, and gene duplications, has offered numerous challenges to developing a genotyping strategy. We describe a comprehensive CYP2D6 genotyping strategy that employs both a PCR/Invader genotyping assay system and an Invader genomic copy number assay The Invader system is a homogeneous, isothermal, highly specific, and robust signal amplification system. Resultsfrom II CYP2D6 assays in an alle frequency study compare well to published allele frequency values for Caucasians. Further, Invader assays provided unambiguous genotyping determinations for 100% of the 171 samples that yielded a visible PCR product on an agarose gel. A copy number assay yielded only one equivocal result in 205 samples. We identified 17 single-copy individuals and 17 three-copy (or more) individuals.
authors.affiliation: Third Wave Technologies, Madison, WI 53719-1256, USA.
authors.forename: Matt
authors.initials: M
authors.lastname: Nevilie
authors.forename: Rebecca
authors.initials: R
authors.lastname: Selzer
authors.forename: Brian
authors.initials: B
authors.lastname: Aizenstein
authors.forename: Megan
authors.initials: M
authors.lastname: Maguire
authors.forename: Kirk
authors.initials: K
authors.lastname: Hogan
authors.forename: Robert
authors.initials: R
authors.lastname: Walton
authors.forename: Ken
authors.initials: K
authors.lastname: Welsh
authors.forename: Bruce
authors.initials: B
authors.lastname: Neri
authors.forename: Monika
authors.initials: M
authors.lastname: de Arruda
mesh_terms: D000483:Alleles; D001483:Base Sequence; D019389:Cytochrome P-450 CYP2D6 / Q000235:genetics*; D004252:DNA Mutational Analysis / Q000295:instrumentation* / Q000379:methods*; D017931:DNA Primers; D015342:DNA Probes; D005189:False Positive Reactions; D005787:Gene Frequency; D015894:Genome, Human; D005838:Genotype; D006801:Humans; D008969:Molecular Sequence Data; D010641:Phenotype; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D020641:Polymorphism, Single Nucleotide; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA / Q000295:instrumentation / Q000379:methods; D017385:Sequence Homology
chemical_list: D017931:DNA Primers; D015342:DNA Probes; D019389:Cytochrome P-450 CYP2D6