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  • Title: Determination of CYP2D6 gene copy number by multiplex polymerase chain reaction analysis.
    Author: Leandro-García LJ, Leskelä S, Montero-Conde C, Landa I, López-Jimenez E, Letón R, Seeringer A, Kirchheiner J, Cascón A, Robledo M, Rodríguez-Antona C.
    Journal: Anal Biochem; 2009 Jun 01; 389(1):74-6. PubMed ID: 19303860.
    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.
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