Determination of CYP2D6 gene copy number by multiplex polymerase chain reaction analysis.
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.
Leandro-García
Luis J
LJ
Hereditary Endocrine Cancer Group, Spanish National Cancer Research Center (CNIO), 28029 Madrid, Spain.
Leskelä
Susanna
S
Montero-Conde
Cristina
C
Landa
Iñigo
I
López-Jimenez
Elena
E
Letón
Rocío
R
Seeringer
Angela
A
Kirchheiner
Julia
J
Cascón
Alberto
A
Robledo
Mercedes
M
Rodríguez-Antona
Cristina
C
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
D019389:Cytochrome P-450 CYP2D6