序列特异性引物多聚酶链反应方法进行大样本多基因的多态性鉴定

摘要

Objective The goal of this experiment was to aid clinical diagnoses of diseases by optimizing the sequence-specific primers-PCR (PCR-SSP) procedure to identify polymorphisms for multiple genes simultaneously.Methods The PCR-SSP method,that was optimized in our laboratory,was used to analyze polymorphisms of the following genes:mutations of -308A/G and -238G/A in TNF-α,-174G/C in IL-6,C/T mutation at 188 in Exton f CYP2D6 *10B.Results Polymorphism analysis for multiple genes and many clinical samples can be performed simultaneously with one PCR program,showing clear genotype and quick and accurate genotyping.Conclusion The optimized PCR-SSP is suitable for polymorphism analysis of the large-sample polygenic mononucleotide point mutation,and it can be widely applied in clinical test for low cost,quickness and accuracy.%目的 建立多个基因同时进行多态性分析的序列特异性引物多聚酶链反应(PCR-SSP)方法,以满足其在临床检验中进行基因多态性鉴定时的应用.方法 应用优化后的PCR-SSP的方法分析以下基因的多态性:TNF-α-308A/G和-238G/A 变异、IL-6 -174G/C 变异、CYP2D6*10B外显子第188位的C/T变异.结果 应用同一个PCR扩增程序,可同时对多个基因、多个临床样本的基因多态性同时进行分析,基因型清晰,而且基因分型快速、准确.结论 优化后的PCR-SSP适合对大样本、多个基因的单核苷酸位点变异进行多态性分析,值得在临床检验的应用中推广.