Department of Pharmacology and Physiology, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Pathumwan, Bangkok 10330, Thailand.
Division of Molecular Genetics, Department of Research and Development, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkoknoi, Bangkok 10700, Thailand.
Clin Biochem. 2011 Sep;44(13):1144-1152. doi: 10.1016/j.clinbiochem.2011.06.985. Epub 2011 Jul 6.
To develop CYP2D6 genotyping scheme for accurate allele calling and reliable estimation of functional allele dosage in Thais.
We analyzed CYP2D6 copy numbers by pentaplex PCR coupled with semi-quantitative denaturing high performance liquid chromatography (DHPLC)-based technique. Ten common SNPs were genotyped from CYP2D6 gene product using single base extension (SBE) followed by DHPLC analysis. This detection scheme was compared with real-time PCR and conventional PCR-RFLP for cost-effectiveness.
The distribution of CYP2D6 gene copy numbers in our population ranged from zero (0.69%), one (7.99%), two (60.07%), three (28.13%) and four (3.13%). The most commonly detected SNPs were related to CYP2D610 haplotype. CYP2D636 in tandem with CYP2D6*10B is the major rearrangement type in Thais (18.75%).
Multiplex PCR coupled with DHPLC-based strategy is convenient and reliable method for CYP2D6 genotyping offering sufficient allele coverage for Asians. Both cost and analytical time saving were shown and the method could potentially be modified to accommodate CYP2D6 genotyping in other ethnics.
制定 CYP2D6 基因分型方案，以准确进行等位基因分型和可靠估计泰国人群中功能性等位基因剂量。
我们通过五重 PCR 结合半定量变性高效液相色谱 (DHPLC) 技术分析 CYP2D6 拷贝数。使用单碱基延伸 (SBE) 结合 DHPLC 分析从 CYP2D6 基因产物中对 10 个常见 SNP 进行基因分型。该检测方案与实时 PCR 和常规 PCR-RFLP 进行成本效益比较。
我们人群中 CYP2D6 基因拷贝数的分布范围从 0（0.69%）、1（7.99%）、2（60.07%）、3（28.13%）和 4（3.13%）。最常见的 SNP 与 CYP2D610 单倍型相关。CYP2D636 与 CYP2D6*10B 串联是泰国的主要重排类型（18.75%）。
五重 PCR 结合 DHPLC 策略是一种方便可靠的 CYP2D6 基因分型方法，为亚洲人提供了足够的等位基因覆盖。显示出成本和分析时间的节省，并且该方法可以进行修改以适应其他种族的 CYP2D6 基因分型。
