[用于CYP2C9*3（1075A/C，rs1057910）和VKORC1（-1639A/G，rs9923231）基因分型的高分辨率熔解方法的开发]

[Development of the high resolution melting method for genotyping CYP2C9*3 (1075A/C, rs1057910) and VKORC1 (-1639A/G, rs9923231)].

作者信息

Cui Guang-lin, Ding Hu, Xu Yu-jun, Chen Chen, Wang Dao-wen

机构信息

Department of Cardiology, Institute of Hypertension, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.

出版信息

Zhonghua Xin Xue Guan Bing Za Zhi. 2012 Jun;40(6):477-81.

PMID:22943641

Abstract

OBJECTIVE

To establish the high-resolution melting curve (HRM) approach for genotyping CYP2C9*3 (1075A/C, rs1057910) and VKORC1 (-1639A/G, rs9923231) and explore its value on estimation of the Warfarin initial dose in comparison with various traditional genotyping methods.

METHODS

CYP2C9*3 (1075A/C, rs1057910) and VKORC1 (-1639A/G, rs9923231) genotyping was detected in 100 patients receiving Warfarin therapy by the newly developed HRM method and traditional genotyping methods including PCR-restriction fragment length polymorphism (PCR-RFLP), TaqMan probe and DNA sequencing.

RESULTS

The results of the genotypes obtained from above mentioned methods to detect CYP2C9*3 (1075A/C, rs1057910) and VKORC1 (-1639A/G, rs9923231) were similar and consistent. The HRM method is simpler, more economical, and faster compared to the traditional methods. The frequencies of the VKORC1-1639 AA, AG, GG genotypes from these 100 clinical samples were 73 (73%), 23 (23%), 4 (4%), respectively; Frequencies of the CYP2C9 1075 AA, AC, CC genotypes were 94(94%), 6 cases (6%), 0, respectively.

CONCLUSIONS

HRM approach can effectively detect CYP2C93 (1075A/C, rs1057910) and VKORC1 (-1639A/G, rs9923231) polymorphisms and this method is simpler, more economical, and faster compared to the traditional methods for detecting CYP2C93 (1075A/C, rs1057910) and VKORC1 (-1639A/G, rs9923231) polymorphisms.

摘要

目的

建立用于细胞色素P450 2C9*3（1075A/C，rs1057910）和维生素K环氧化物还原酶复合体1（VKORC1）（-1639A/G，rs9923231）基因分型的高分辨率熔解曲线（HRM）方法，并与各种传统基因分型方法比较，探讨其在华法林初始剂量估算中的价值。

方法

采用新开发的HRM方法及包括聚合酶链反应-限制性片段长度多态性（PCR-RFLP）、TaqMan探针和DNA测序在内的传统基因分型方法，对100例接受华法林治疗的患者进行细胞色素P450 2C9*3（1075A/C，rs1057910）和VKORC1（-1639A/G，rs9923231）基因分型检测。

结果

上述检测细胞色素P450 2C9*3（1075A/C，rs1057910）和VKORC1（-1639A/G，rs9923231）的方法所获得的基因型结果相似且一致。与传统方法相比，HRM方法更简便、经济且快速。这100例临床样本中VKORC1 -1639 AA、AG、GG基因型的频率分别为73（73%）、23（23%）、4（4%）；细胞色素P450 2C9 1075 AA、AC、CC基因型的频率分别为94（94%）、6例（6%）、0。

结论

HRM方法可有效检测细胞色素P450 2C93（1075A/C，rs1057910）和VKORC1（-1639A/G，rs9923231）多态性，且与检测细胞色素P450 2C93（1075A/C，rs1057910）和VKORC1（-1639A/G，rs9923231）多态性的传统方法相比，该方法更简便、经济且快速。