150例广东籍汉族人肿瘤坏死因子TNFα-863C/A基因突变频率的检测

Detection of mutation frequencies in the TNFα-863 C/A gene with PCR-RFLP in 150 health individuals Guangdong Han population

目的建立一种简便、准确、实用的人肿瘤坏死因子(TNF)α-863 C/A基因突变频率的检测方法 ,并了解广东籍汉族人TNFα-863 C/A C基因的分布特点。方法应用聚合酶链反应(特异性扩增人TNFα-863 C/A基因序列,扩增产物用限制性内切酶以Tai I酶切,琼脂糖凝胶电泳后,观察酶切位点的限制性片段长度多态性图谱,对150例广东籍正常人TNFα-863 C/A基因进行检测,并结合文献进行了不同地区间的分析比较。结果 TNFα-863 C/A C/C野生型纯合子频率为75.33%,C/A杂合子频率为24.00%,A/A突变型纯合子频率为0.67%;突变等位基因频率为0.1267。结论该方法简便、快速、准确,适合于一般实验室检测及大规模的人群调查,TNFα-863 C/A基因多态性在不同地区间分布存在着一定的差异。

Objective To establish a simple,accurate and practical method for TNFα-863 C/A genotyping and identify the distribution feature of TNFα-863 C/A gene in the Hans in Chinese.Methods TNFα-863 C/A gene sequences were amplified by using polymerase chain reaction（PCR）.The PCR products were digested with Tai I and subjected to electrophoresis on agarose gels.The patterns of restriction fragment length polymorphism（RFLP） of TNFα-863 C/A gene were distinguished.Results 150 health individuals were investigated with this PCR-RFLP method.The gene frequency of TNFα-863 C/A was as followed： TNFα-863 C/A C/C,C/A,A/A genotype frequency： 75.33%,24.00%,0.67%,respectively;TNFα-863 C,TNFα-863 A genefrequency： 0.8733 and 0.1267.Conclusion The PCR-RFLP method was considered a rapid and simple technique for obtaining accurate results and also suitable for routine laboratories and large-scale population studies,TNFα-863 C/A genes distribution in the Hans in Chinese is undifferent from those in other areas.