扩增引进限制性酶切位点技术对两种CYP21基因突变的检测  

Technique of PCR-ACRS for the detection of CYP21 gene mutations

作　　者：廖相云[1] 张雅芬[1] 顾学范[1] 

机构地区：[1]上海第二医科大学附属新华医院,上海市儿科医学研究所200092

出　　处：《中华医学遗传学杂志》2003年第5期449-451,共3页Chinese Journal of Medical Genetics

基　　金：国家自然科学基金(39870 778);上海市高教局基金( 0 1 B0 4 )

摘　　要：目的　建立一种快速、可靠的 CYP2 1基因突变的检测方法。方法　收集 50例 2 1-羟化酶缺乏症患者及部分先证者家系的外周血 ,并提取 DNA。用计算机软件 (DNAssist)对 CYP2 1基因突变相应的限制性内切酶的酶切位点进行分析 ,使用 PCR结合扩增引进限制性酶切位点的方法对没有酶切位点改变的 I172 N和 R3 56W两种 CYP2 1基因突变进行检测。结果　有 2 1例患者发现 I172 N突变 ,其中 3例纯合子 ,18例杂合子 ;8例患者发现 R3 56W突变 ,皆为杂合子表现 ,其发生频率与文献报道的相似。并且通过I172 N和 R3 56W两种突变的各 5个家系分析 ,皆符合常染色体隐性遗传的特点。结论　以 PCR为基础的扩增引进限制性酶切位点法简便、可靠 ,是快速检测 CYP2Objective: To establish a rapid method of detecting CYP21 gene mutations. Methods: Fifty Chinese patients with 21-hydroxylase deficiency and some of their families were investigated. Blood samples were obtained for extraction of peripheral blood lymphocytes. A search for restriction sites discriminating between the morbid and the normal in CYP21 gene was made by the computer program DNAssist. PCR-based amplication-created restriction site (PCR-ACRS) was performed at I172N and R356W which are not natural recognition sequence. In addition, I172N and R356W were analysed in five families which conform to the applicability of PCR-ACRS. Results: In 50 identified 21-hydroxylase deficient Chinese patients, 21 were found to have I172N (3 were homozygote, 18 were heterozygote); 8 were found to have R356W, all of them were heterozygote. By analysing the families, the findings were consistent with the characteristics of autosomal recessive genetic deficiency. Conclusion: Analysis of CYP21 gene point mutations using PCR-ACRS is relatively simple, accurate and feasible.

关 键 词：CYP21 基因突变 2l-羟化酶 先天性肾上腺皮质增生症 CAH 聚合酶链反应 PCR 

分 类 号：R346[医药卫生—基础医学]