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J Biomed Biotechnol. 2005;2005(3):248-53.

Determination of cytochrome P450 2D6 (CYP2D6) gene copy number by real-time quantitative PCR.

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1
INSERM UMRS 490, Université René Descartes - Paris 5, 45 rue des Saints-Péres, 75270 Paris, France.

Abstract

Gene dosage by real-time quantitative PCR has proved to be accurate for measuring gene copy number. The aim of this study was to apply this approach to the CYP2D6 gene to allow for rapid identification of poor and ultrarapid metabolizers (0, 1, or more than 2 gene copy number). Using the 2(-Delta Delta Ct) calculation method and a duplex reaction, the number of CYP2D6 gene copies was determined. Quantitative PCR was performed on 43 samples previously analyzed by Southern blotting and long PCR including 20 samples with a heterozygous deletion, 11 with normal copy number (2 copies), and 12 samples with duplicated genes. The average ratio ranged from 1.02 to 1.28, 1.85 to 2.21, and 2.55 to 3.30, respectively, for the samples with 1 copy, 2 copies, and 3 copies. This study shows that this method is sensitive enough to detect either a heterozygous gene deletion or duplication.

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