Rapid Single-Nucleotide Polymorphism Detection of Cytochrome P450 (CYP2C9) and Vitamin K Epoxide Reductase (VKORC1) Genes for the Warfarin Dose Adjustment by the SMart-Amplification Process Version 2
Rapid Single-Nucleotide Polymorphism Detection of Cytochrome P450 (CYP2C9) and Vitamin K Epoxide Reductase (VKORC1) Genes for the Warfarin Dose Adjustment by the SMart-Amplification Process Version 2
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DOI：
10.1373/clinchem.2008.115295
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发表时间：
2009-04-01
影响因子：
9.3
通讯作者：
Horiuchi, Ryuya
中科院分区：
文献类型：
--
作者：
Aomori, Tohru;Yamamoto, Koujirou;Horiuchi, Ryuya
背景技术背景：CYP 2C 9（细胞色素P450，家族2，亚家族C，多肽9）基因（CYP 2C 9 *2，CYP 2C 9 *3）和VKORC 1（维生素K环氧化物还原酶复合物，亚基1）基因（-1639G>A）的多态性极大地影响药物华法林的维持剂量。在开药前对患者的基因型进行预筛选有助于更快地个体化确定适当的维持剂量，最大限度地降低不良反应和血栓栓塞事件复发的风险。采用目前的方法，如果基因分型是为了确定负荷剂量，治疗可能会延迟几个小时至1天。方法：我们开发了一种单核苷酸多态性（SNP）检测分析的基础上的SMART扩增过程版本2（SMAP 2）分析CYP 2C 9 *2，CYP 2C 9 -3，和VKORC 1 - 1639 G>A多态性。从同意的参与者的血液被直接用于在一个封闭的管实时检测没有DNA纯化后1小时内获得结果bloodcollection.RESULTS：我们分析了125血液样本SMAP 2和PCR-RFLP方法。结果显示完美的concordance.CONCLUSIONS：结果验证了SMAP 2的准确性确定的SNPs的个性化华法林治疗的关键。SMAP 2提供了速度，样品制备的简单性，等温扩增的便利性和分析设计的灵活性，这些都是传统基因分型技术的显着优势。在这个例子和其他临床情况下，需要进行基因检测以立即做出更明智的治疗决定，基于SMAP 2的诊断具有关键优势。(C)2008年美国临床化学协会
BACKGROUND: Polymorphisms of the CYP2C9 (cytochrome P450, family 2, subfamily C, polypeptide 9) gene (CYP2C9*2, CYP2C9*3) and the VKORC1 (vitamin K epoxide reductase complex, subunit 1) gene (-1639G>A) greatly impact the maintenance dose for the drug warfarin. Prescreening patients for their genotypes before prescribing the drug facilitates a faster individualized determination of the proper maintenance dose, minimizing the risk for adverse reaction and re-occurrence of thromboembolic episodes. With current methodologies, therapy can be delayed by several hours to 1 day if genotyping is to determine the loading dose. A simpler and more rapid genotyping method is required.METHODS: We developed a single-nucleotide polymorphism (SNP)-detection assay based on the SMart Amplification Process version 2 (SMAP 2) to analyze CYP2C9*2, CYP2C9-3, and VKORC1 -1639G>A polymorphisms. Blood from consenting participants was used directly in a closed-tube real-time assay without DNA purification to obtain results within I h after blood collection.RESULTS: We analyzed 125 blood samples by both SMAP 2 and PCR-RFLP methods. The results showed perfect concordance.CONCLUSIONS: The results validate the accuracy of the SMAP 2 for determination of SNPs critical to personalized warfarin therapy. SMAP 2 offers speed, simplicity of sample preparation, the convenience of isothermal amplification, and assay-design flexibility, which are significant advantages over conventional genotyping technologies. In this example and other clinical scenarios in which genetic testing is required for immediate and better-informed therapeutic decisions, SMAP 2-based diagnostics have key advantages. (C) 2008 American Association for Clinical Chemistry