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(英文)

Large-sample and polygenic identification of polymorphisms using sequence-specific primers-PCR

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。

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 polymorphlsms 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.