荧光PCR-熔解曲线法检测新疆回族人群CYP2C9和VKORC1基因多态性研究

目的应用荧光PCR-熔解曲线法检测新疆回族人群细胞色素P450酶2C9(CYP2C9)和维生素K环氧化物还原酶复合体亚单位1(VKORC1)基因多态性,研究新疆回族人群CYP2C9和VKORC1基因分布及基因突变频率情况,并评价荧光PCR-熔解曲线法临床使用的适用性。方法采用荧光PCR-熔解曲线法和测序法对比检测CYP2C9*2、CYP2C9*3和VKORC1(-1639G/A)的基因多态性。结果 228例新疆回族人群中,共检测出CYP2C9*1/*1标本199例,CYP2C9*1/*2标本2例,CYP2C9*1/*3标本26例,CYP2C9*3/*3标本1例,未检测出CYP2C9*2/*2和CYP2C9*2/*3标本。VKORC1(-1639G/A)检测出2种等位基因G和A,其中VKORC1-1639G/G型标本检出2例,VKORC1-1639G/A型标本检出39例,VKORC1-1639A/A型标本检出187例,与测序法对比检测比较,荧光PCR-熔解曲线法与测序法检测结果完全一致。结论新疆回族人群同样具有CYP2C9基因*2、*3位点和VKORC1基因-1639位点的多态性,其发生频率与新疆维吾尔族及其他地域人群的具有一定差异,所采用的荧光PCR-熔解曲线法用于基因多态性检测可满足临床检测要求。

Study on fluorescence PCR-melting curve method for detecting CYP2C9 and VKORC1 gene polymorphism in Xinjiang Hui population

Objective To use the fluorescence PCR‐melting curve method to detect CYP2C9 and VKORC1 gene polymorphism in Xinjiang Hui population ,to analyze their gene distribution and gene mutation frequency ,and to evaluate the clinical applicability of the fluorescence PCR‐melting curve method .Methods The fluorescence PCR‐melting curve method and sequencing method were adopted to contrastively detect CYP2C9*2 ,CYP2C9*3 and VKORC1(‐1639G/A)gene polymorphism .Results Among detected 228 Xinjiang Hui individuals ,199 cases of CYP2C9*1/*1 ,2 cases of CYP2C9*1/*2 ,26 cases of CYP2C9*1/*3 and only 1 case of CYP2C9*3/*3 were detected ,no case of CYP2C9*2/*2 and CYP2C9*2/*3 was detected .Two kinds of allele G and A were detected for VKORC1(‐1639G/A) ,in which VKORC1‐1639G/G type was detected in 2 cases ,VKORC1‐1639G/A type was detected in 39 cases and VKORC1‐1639A/A type was detected in187 cases ,compared with the sequencing method ,the results of the fluorescence PCR‐melting curve method were completely consistent .Conclusion Xinjiang Hui population also has CYP2C9 gene *2 ,*3 loci and VKORC1 gene(‐1639G/A) locus polymorphism ,their occurrence frequency has a certain difference with Xingjiang Uygur and other regional populations ,the adopted fluorescence PCR‐melting curve method used in the gene polymorphism detection can meet clinical detection requirements .