四引物突变特异性扩增系统法检测巨噬细胞移动抑制因子基因-173位点单核苷酸多态性分布

Distribution of MIF -173 single nucleotide polymorphism checked by tetra-primer amplification refractory mutation system

目的探讨中国浙江地区汉族人群巨噬细胞移动抑制因子(macrophagemigrationinhibitoryfac tor,MIF)基因-173位点单核苷酸多态性(singlenucleotidepolymorphism,SNP)分布。方法收集浙江地区142名无血缘关系健康个体的静脉血,提取DNA,分别应用四引物突变特异性扩增系统(amplificationrefracto rymutationsysntem,ARMS)法和限制性片段长度多态性PCR方法对MIF基因-173位点SNP多态性进行分型,并将PCR产物克隆及测序鉴定。结果MIF基因-173位点检测到3种基因型,其基因型分布皆符合Hardy Weinberg平衡定律。四引物ARMS法和限制性片段长度多态性PCR两种方法结果完全一致。统计分析显示,中国汉族人MIF基因-173位点等位基因和基因型频率分布与欧洲白人差异有统计学意义(P<0.01),与日本人群的差异无统计学意义(P>0.05)。结论四引物ARMS法是一种准确、快速和经济的SNP测定方法。MIF基因-173位点等位基因频率分布具有种族的差异性。

Objective To study the distribution of macrophage migration inhibitory factor gene （MIF） - 173 single nucleotide polymorphism （SNP） of Chinese Han population in Zhejiang province. Methods The DNA samples were extracted from EDTA-bloed of 142 unrelated healthy individuals. Alleles of MIF - 173 SNP were genotyped by using the techniques of tetra-primer amplification refractory mutation system （ARMS） and restriction fragment length polymorphisms （RFLP）-PCR. Meantime the PCR products were cloned and sequenced. Results The authors detected three kinds of genotypes at the MIF - 173 locus, and no deviation was observed from Hardy-Weinberg equilibrium. The final results were the same completely, whatever either tetra-primer ARMS or RFLP-PCR was used to check the MIF - 173 single nucleotide polymorphism. Statistical analysis showed that the distributions of MIF - 173 SNP alleles and genotype frequencies were significantly different between Chinese Hart population and European Caucasian（ P 〈 0.01 ）, but no significant difference demonstrated to happen between Chinese and Japanese（ P 〉 0.05）. Condusion Tetra-primer ARMS is a accurate, rapid and economical method for SNP genotyping.There exists ethnic difference in the distribution of MIF - 173 SNP alleles.