摘要
目的:建立等位基因特异性PCR检测脂联素基因与2型糖尿病相关SNP位点的方法。方法:针对脂联素基因G531A,G545C,G11963T,T12194G和T13320C5个SNP位点设计野生型和突变型特异性引物,第一轮PCR的上游引物或下游引物与等位基因特异性引物进行半巢式PCR。在同一SNP位点如果野生型引物出现扩增产物而突变型引物未出现扩增产物,则判定该位点为野生型,反之,为突变型;如两引物均出现扩增产物则为杂合子。DNA测序证实等位基因特异性PCR检测结果。结果:该等位基因特异性PCR检测方法对研究样本均有扩增,且每个SNP位点均检出相应的野生型、突变型和混合型个体。测序结果与等位基因特异性PCR检测结果吻合。结论:建立的等位基因特异性PCR检测脂联素基因SNP的方法灵敏可靠,可快速检测与2型糖尿病相关的5个SNP位点。
Objective： To establish an allele specific PCR method for detecting adiponectin gene SNPs related to type 2 diabetes.Methods： Primers,which could specifically amplify wild types or their mutants of 5 SNPs,G531A,G545C,G11963T,T12194G and T13320C in adiponectin gene were designed.A semi-nest allele PCR was carried out with the specific upper primer or lower primer.If wild type primer amplification happened while mutant primer amplification did not,the SNP point was considered as wild type.If mutant primer amplification happened while wild type primer amplification did not,the SNP point was considered as mutant.And if both wild type and mutant amplification happened,the SNP point was considered as heterozygote.The results of allele specific PCR were confirmed by sequencing.Results： Specific amplifications were obtained with all of the allele specific primers.Wild type,mutant and heterozygote were detected in the 5 SNP points.Conclusions： This allele specific PCR method is effective,specific and rapid in detecting the 5 SNPs of adiponectin gene related to type 2 diabetes.
出处
《华夏医学》
CAS
2011年第4期403-406,共4页
Acta Medicinae Sinica
基金
广西留学回国人员基金项目(桂科回0448024)
