聚合酶链反应-序列特异性引物方法检测儿科疾病基因多态性

Polymerase Chain Reaction-Sequence Special Primers in Detecting Gene Polymorphism of Pediatrics Diseases

目的建立多个基因多态性分型的聚合酶链反应-序列特异性引物(PCR-SSP)的方法,以推广其在儿科疾病分子水平研究中的应用。方法应用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。结果应用同一个PCR扩增程序,可同时对多个基因、多个临床样本的多态性进行分析,基因型清晰,且基因分型快速、准确。结论PCR-SSP适合对大样本、多基因的多态性进行分析,成本低、快速、准确,在儿科疾病分子水平研究中前景良好。

Objective To establish an approach of polylnerase chain reaction sequence special primers（PCR - SSP） to perform polymorphism analysis for multiple genes simultaneously, in order to be used in clinical analysis. Methods The optimized PCR SSP approach was used to analyze the polymorphism of the following genes： mutation of 308A/G and - 238G/A in tumor necrosis factor-α （TNF- α）, - 174G/C in interleukin - 6（ IL - 6）. Results Polymorphism analysis of multiple genes and many clinical samples could be simultaneously performed with one PCR program, showing clear genotype, quick and accurate genotype. ConcIusion The optimized PCR - SSP is suitable for polymorphism analysis of the large-sample polygenic mononcleeotide point mutation,and it can be widely applied in clinical test for low cost, quickness and accuracy.