基质金属蛋白酶9C1562T基因突变频率的检测

目的：建立一种简便、准确、实用的人基质金属蛋白酶9C1562T基因突变频率的检测方法,并了解汉族人基质金属蛋白酶9C1562T基因的分布特点.方法：115例广东籍汉族人血样本取自2004年9月广州医学院卫生所体检的正常人,应用聚合酶链反应（PCR）特异性扩增人基质金属蛋白酶9C1562T基因序列,扩增产物用限制性内切酶SphⅠ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（RFLP）图谱.结果：运用PCR-RFLP法检测了115名广东籍汉族人基质金属蛋白酶9基因C1562T点突变,其中野生型纯合子频率为87.83%,杂合子频率为10.43%,突变型纯合子频率为1.74%.突变等位基因频率为0.0694.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,汉族人基质金属蛋白酶9C1562T基因的分布与其他地区人群的分布无明显差异.

Detection of MMP-9 Mutation Frequencies in Healthy Individuals

Objective:To establish a simple, accurate and practical method for MMP-9 C1562T genotyping and identify the distribution of MMP-9C1562T gene in the Hans in Chinese. Method:115 case in the Hans in Chinese MMP-9 C1562T gene sequences were amplified by using polymerase chain reaction (PCR). The PCR products were digested with SphI and subjected to electrophoresis on agarose gels. The patterns of restriction fragment length polymorphism (RFLP) of MMP-9 C1562T gene were distinguished. Results: 115 healthy individuals were investigated with this PCR-RFLP method. The prevalence of genotypes for normal homozygotes, normal/mutant heterozygotes and mutant homozygotes were 87. 83% , 10.43% and 1.74% respectively. The frequency of mutant allele in the subjects was 0.0694. Conclusion: PCR-RFLP was considered a rapid and simple technique for obtaining accurate results and also suitable for routine laboratory and large-scale population studies. MMP-9 C1562T genes distribution in the Hans in Chinese is not significantly different from those in other areas.