Determination of CYP2D6 gene copy number by multiplex polymerase chain reaction analysis

Luis J Leandro-García; Susanna Leskelä; Cristina Montero-Conde; Iñigo Landa; Elena López-Jimenez; Rocío Letón; Angela Seeringer; Julia Kirchheiner; Alberto Cascón; Mercedes Robledo; Cristina Rodríguez-Antona

doi:10.1016/j.ab.2009.03.021

Determination of CYP2D6 gene copy number by multiplex polymerase chain reaction analysis

Anal Biochem. 2009 Jun 1;389(1):74-6. doi: 10.1016/j.ab.2009.03.021. Epub 2009 Mar 20.

Authors

Luis J Leandro-García¹, Susanna Leskelä, Cristina Montero-Conde, Iñigo Landa, Elena López-Jimenez, Rocío Letón, Angela Seeringer, Julia Kirchheiner, Alberto Cascón, Mercedes Robledo, Cristina Rodríguez-Antona

Affiliation

¹ Hereditary Endocrine Cancer Group, Spanish National Cancer Research Center (CNIO), 28029 Madrid, Spain.

PMID: 19303860
DOI: 10.1016/j.ab.2009.03.021

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cytochrome P-450 CYP2D6 / genetics*
Gene Dosage / genetics*
Genotype
Humans
Polymerase Chain Reaction / methods*
Polymorphism, Restriction Fragment Length / genetics
Polymorphism, Single Nucleotide / genetics

Substances

Cytochrome P-450 CYP2D6