title: Determination of CYP2D6 gene copy number by multiplex polymerase chain reaction analysis.
abstract: Cytochrome P450 2D6 (CYP2D6) copy number variation (CNV) influences the metabolism of 15-25% of clinical drugs. Here we describe a novel multiplex polymerase chain reaction (PCR) analysis method that accurately detects CYP2D6 CNV and CYP2D6*9 allele. It includes the amplification of 2 CYP2D6 and 7 control (AQP1, CYP3A4, MDR1, and SDHB) fluorescent PCR products that are separated on a capillary sequencer and normalized using reference samples. The technique was validated using 27 PCR-restriction fragment length polymorphism (RFLP) pregenotyped samples and further tested in 75 Caucasian samples. The method assigns the correct CYP2D6 copy number, independent of already characterized CYP2D6 single nucleotide polymorphisms (SNPs), and could easily be applied to clinical samples.
authors.affiliation: Hereditary Endocrine Cancer Group, Spanish National Cancer Research Center (CNIO), 28029 Madrid, Spain.
authors.forename: Luis J
authors.initials: LJ
authors.lastname: Leandro-García
authors.forename: Susanna
authors.initials: S
authors.lastname: Leskelä
authors.forename: Cristina
authors.initials: C
authors.lastname: Montero-Conde
authors.forename: Iñigo
authors.initials: I
authors.lastname: Landa
authors.forename: Elena
authors.initials: E
authors.lastname: López-Jimenez
authors.forename: Rocío
authors.initials: R
authors.lastname: Letón
authors.forename: Angela
authors.initials: A
authors.lastname: Seeringer
authors.forename: Julia
authors.initials: J
authors.lastname: Kirchheiner
authors.forename: Alberto
authors.initials: A
authors.lastname: Cascón
authors.forename: Mercedes
authors.initials: M
authors.lastname: Robledo
authors.forename: Cristina
authors.initials: C
authors.lastname: Rodríguez-Antona
mesh_terms: D019389:Cytochrome P-450 CYP2D6 / Q000235:genetics*; D018628:Gene Dosage / Q000235:genetics*; D005838:Genotype; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length / Q000235:genetics; D020641:Polymorphism, Single Nucleotide / Q000235:genetics
chemical_list: D019389:Cytochrome P-450 CYP2D6