肾上腺脑白质营养不良家系ABCD1基因突变研究  

Mutation analysis of the ABCD1 gene in adrenoleukodystrophy pedigrees

作　　者：张林[1,2,3] 林宇翔[1] 兰风华[1] 王志红[1] 

机构地区：[1]福建医科大学福总临床医学院(南京军区福州总医院)临床遗传与实验医学科,福建福州350025 [2]三峡大学人民医院 [3]宜昌市第一人民医院

出　　处：《中国实用妇科与产科杂志》2014年第3期192-196,共5页Chinese Journal of Practical Gynecology and Obstetrics

基　　金：国家自然科学基金青年科学基金项目(31200932);福建省自然科学基金青年创新项目(2012J05158)

摘　　要：目的分析肾上腺脑白质营养不良(adrenoleukodystrophy,ALD)家系的先证者及ALD携带者所孕育胎儿的ABCD1基因突变。方法首先从RNA途径,应用RT-PCR技术,对2010年10月至2012年3月福建医科大学福总临床医学院5个家系先证者或其母亲的ABCD1基因编码区进行序列测定查找突变,再通过基因组DNA途径进行突变验证。产前诊断采用STR位点分析方法排除母体基因组DNA污染后,应用DNA测序或PCR-限制性酶切方法对5个胎儿的羊水基因组DNA进行突变分析。结果在5个家系的先证者或先证者母亲分别发现5个不同位点的ABCD1基因突变,依次为c.796G>A,c.521A>G,c.1523C>T,c.1534G>A,c.1252C>T。产前诊断结果显示:5个胎儿均为男性,胎儿1、2、3、4存在与先证者相同的突变,胎儿5未检测到与先证者5相同的突变。结论 ABCD1基因突变分析是ALD分子诊断及产前分子诊断准确、可靠的方法。Objective To carry out mutation analysis of the ABCD1 gene in 5 adrenoleukodystrophy （ALD） pedigrees. Methods The coding region of ABCD1 gene of patients or their mothers was amplified from total RNA of the peripheral blood leukocytes by RT-PCR. The PCR products were purified and directly sequenced. To confirm the mutations, the genomic DNA of the patients and their family members was analyzed by direct sequencing. Maternal contamination was excluded by fluorescent STR profiling. The genomie DNA of amniotic fluid cells （AFC） of 5 fetuses was analyzed by PCR- restrictive digestion or direct sequencing. Results Five base substitutions （ e. 796G 〉 A, c. 521A 〉 G, c. 1523C 〉 T, c. 1534G 〉 A and e. 1252C 〉T） were identified in five ALD pedigrees, which resulted in five missense mutations （p. Gly266Arg, p. Tyr174Cys, p. Pro508Leu, p. Gly512Ser and p. Arg418Trp）. The 5 fetuses were males. The same mutation as the proband was detected in the ABCD1 gene of fetus 1, fetus 2, fetus 3 and fetus 4. The same mutation as the proband was not detected in fetus 5. Conclusion Mutation analysis of the ABCD1 gene is an efficient and accurate method for molecular diagnosis of ALD.

关 键 词：肾上腺脑白质营养不良 分子诊断 产前分子诊断 ABCD1基因 基因突变 

分 类 号：R714.2[医药卫生—妇产科学]