Cui Guang-lin, Ding Hu, Xu Yu-jun, Chen Chen, Wang Dao-wen
Department of Cardiology, Institute of Hypertension, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.
Zhonghua Xin Xue Guan Bing Za Zhi. 2012 Jun;40(6):477-81.
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.
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.
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.
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.
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。
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。
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。
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。
