动脉粥样硬化患者CETP基因两种主要突变型的检测

目的 :检测中国人群动脉粥样硬化 ( AS)患者和正常人的胆固醇酯转运蛋白 ( CETP)基因 15外显子 D44 2 G突变和 14内含子 I14A突变。　方法 :应用聚合酶链反应 -限制性内切酶片段长度多态性分析 ( PCR- RFL P)的方法检测 110例脑梗死患者和 87例心肌梗死患者胆固醇酯转运蛋白基因 D44 2 G突变和 I14A突变。首先用 PCR分别扩增包含人 CETP基因 15外显子第 15 0 6位碱基和 14内含子 1位碱基的基因序列 ,扩增产物分别用限制性内切酶 Msp I和 Nde I酶切 ,聚丙烯酰胺凝胶电泳后分析结果。　结果 :110例脑梗死患者中发现 4例 D44 2 G杂合子 ,1例 I14A杂合子 ,突变率分别为 3.6 %、0 .91% ;87例心肌梗死患者中发现 3例 D44 2 G杂合子 ,1例 I14A杂合子 ,突变率分别为 3.45 %、1.0 5 % ;10 0例正常人中发现 4例 D44 2 G杂合子 ,1例 D44 2 G纯合子 ,1例 I14A杂合子 ,突变率分别为 5 %、1% ,两病例组与对照组比较差异不明显 ( P>0 .0 5 )。　结论 :正常个体和动脉粥样硬化患者CETP基因 D44 2 G突变和 I14A突变的发生率无差异

Detection of exon 15 missense mutations(D442G) and intron 14 splice defect(I14A) in cholesteryl ester transfer protein gene in atherosclerosis

Objectives: To examine the frequency of exon 15 missense mutation (D442G) and intron 14 splice defect (I14A) in cholesteryl ester transfer protein gene in Chinese atherosclerotic subjects. Methods: Using PCR RFLP to develop a simple and practical method for detecting two prevalent mutations in CETP gene. The amplified DNA containing the base position of exon 15 1506, was digested with MspI and the other containing the intron 14 1 base was digested with NdeI. Then the products were subjected to electrophoresis in polyacrylamide gel. After staining in ethidium bromide, DNA fragments were visualized on a standard UV transilluminator. Results: We screened 110 patients with cerebral infarction and found 4 heterozygotes (3.6%) with CETP gene D442G mutation and 1 heterozygote (0.91%) with I14A mutation. In 87 MIS we found 3 heterozygotes (3.45%) and 1 heterozygote (1.05%) respectively. There was no significant difference in patients and controls (5% and 1% respectively). Conclusions: The frequency of CETP gene defect in atherosclerosis and normal individuals was not different significantly (P> 0.05).