白细胞介素6基因启动子区两个突变位点多态性检测方法的建立

Establishment of simultaneous rapid genotyping of two mutation sites in the promoter region of interleukin 6 gene

目的建立一种快速检测 IL-6基因启动子区-597G/A、-572G/C 多态性的双重实时荧光 PCR 方法。方法采用1对引物,2对荧光标记探针,结合荧光共振能量转移原理和熔点曲线分析技术,检测 IL-6基因启动子区-572G/C,-597G/A 2个位点多态性。结果用建立的双重实时荧光PCR 方法对123名健康查体者进行检测,发现中国汉族人 IL-6基因启动子^-572位点有3种基因型,分别为 GG,GC,CC 型;-597位点仅发现4名为 GA 型,其余均为 GG 型,尚未发现 AA 型。结论双重实时荧光 PCR 法简便快速,与其他方法比较具有准确、经济的特点,适合临床基因快速分型。

Objective To establish a rapid assay for genotyping of IL-6 -597G/A and -572G/C polymorphisms by duplex real-time PCR assay. Methods One pair of primers and two pairs of fluorescent hybridization probes had been used to genotype two mutation sites in the promoter region of IL-6 gene by fluorescent resonance energy transfer and melting curves. Results Duplex real-time PCR method for genotyping of two mutation sites simultaneously had been developed and 123 health people samples were analyzed by this new method. The results showed that three genotypes were found in IL-6 gene -572G/C polymorphisms. They were GG, GC and CC genotypes. IL-6 gene promoter -597G/A polymorphism analysis showed 4 cases displayed GA genotype and other possessed GG genotype. No AA genotype had been found. Conclusions Duplex real-time PCR method is simple and fast. It provided an accurate and economic method which is suitable for clinical gene genotyping.