通过多重聚合酶链反应分析测定CYP2D6基因拷贝数。
Determination of CYP2D6 gene copy number by multiplex polymerase chain reaction analysis.
作者信息
Leandro-García Luis J, Leskelä Susanna, Montero-Conde Cristina, Landa Iñigo, López-Jimenez Elena, Letón Rocío, Seeringer Angela, Kirchheiner Julia, Cascón Alberto, Robledo Mercedes, Rodríguez-Antona Cristina
机构信息
Hereditary Endocrine Cancer Group, Spanish National Cancer Research Center (CNIO), 28029 Madrid, Spain.
出版信息
Anal Biochem. 2009 Jun 1;389(1):74-6. doi: 10.1016/j.ab.2009.03.021. Epub 2009 Mar 20.
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.
细胞色素P450 2D6（CYP2D6）拷贝数变异（CNV）影响15%至25%临床药物的代谢。在此，我们描述了一种新型多重聚合酶链反应（PCR）分析方法，该方法可准确检测CYP2D6 CNV和CYP2D6*9等位基因。它包括扩增2个CYP2D6和7个对照（水通道蛋白1、CYP3A4、多药耐药蛋白1和琥珀酸脱氢酶亚基B）荧光PCR产物，这些产物在毛细管测序仪上分离并用参考样本进行标准化。该技术使用27个经PCR-限制性片段长度多态性（RFLP）预基因分型的样本进行了验证，并在75个白种人样本中进一步测试。该方法能够正确确定CYP2D6拷贝数，与已鉴定的CYP2D6单核苷酸多态性（SNP）无关，并且可以轻松应用于临床样本。
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