等位基因3′末端碱基特异性引物定量PCR检测人脂联素基因单核苷酸多态性  

Single nucleotide polymorphism of human APM1 detected by using allelic 3′-terminal specific primers real-time PCR

作　　者：宋现让[1] 王淑芳[2] 迟伟玲[3] 

机构地区：[1]山东省肿瘤医院基础研究中心,山东济南250117 [2]济南市中心医院,山东济南250014 [3]山东省千佛山医院,山东济南250014

出　　处：《基础医学与临床》2007年第9期1025-1028,共4页Basic and Clinical Medicine

摘　　要：目的建立非探针标记荧光定量PCR的SNP检测技术,分析人脂联素(APM1)基因单核苷酸多态性(SNP)与Ⅱ型糖尿病(T2DM)的关系。方法设计3′末端碱基特异性引物,进行定量PCR反应,通过比较各对引物的扩增效率判断基因型,并进行测序验证。利用该方法确定20例T2DM、24例肥胖及28例同年龄组正常人APM1基因的45位和276位碱基的SNP。结果该方法能有效地区分不同的基因型,与测序结果符合率100%。APM1基因45位SNP与T2DM相关(P<0.05),基因型为TT纯合子者发生T2DM的危险性是G/T和GG者的3倍;而肥胖与APM1基因45和276位点SNP无关。结论等位基因3′末端碱基特异性引物定量PCR可以用于基因SNP检测。APM1基因45位碱基的SNP与T2DM发生有关。Objective To develop an analyzing technique of single nucleotide polymorphism（SNP） based on non- probe real-time quantitative PCR, and to explore the relationship between SNP in human adiponectin （APM1） gene and type 2 diabetes （T2DM）. Methods Design primers with specific 3＇-terminal nucleotides and perform Real- time PCR using SYBR Green dye. Genotypes were determined by comparing the amplification efficiency of each pair of primer and verified by sequencing. SNPs 45 and 276 in APM1 of 20 T2DM, 24 obesity and 28 healthy persons were analyzed. Results This technique could discriminate genotype as efficient as sequencing. The genotype of APM1 at ＋45 site was significantly correlated with T2DM （P 〈0.05 ）. Moreover, the relative risk of T2DM incidence in persons with TF genotype was 3 time more than those with GG and GT. On the contrary, obesity was not correlated with SNP at ＋ 45 nor ＋ 276 sites of APM1 gene. Conclusion Allelic 3＇-terminal specific primers real-time PCR could be used to determine SNPs. SNP at ＋45 sites of APM1 gene correlated with T2DM.

关 键 词：人脂联素基因 单核苷酸多态性 荧光定量PCR 

分 类 号：R450[医药卫生—治疗学]