脑血管狭窄患者Nf1全基因的DHPLC分析(附5例报告)  

DHPLC analysis on Nf1 gene in five patients with cerebrovascular stenosis

作　　者：梁建涛[1] 贾冰冰 邹俊华 霍丽蓉[2] 

机构地区：[1]首都医科大学宣武医院神经外科,北京100053 [2]首都医科大学附属复兴医院神经病学系,中心实验室 [3]北京大学医学部基础医学院医学遗传系

出　　处：《北京医学》2017年第10期991-994,F0002,共5页Beijing Medical Journal

基　　金：国家自然科学基金主任基金(81341036);北京市卫生系统高层次卫生技术人才培养计划"学科骨干"(2013-3-096);北京市西城区优秀人才项目(20160031)

摘　　要：目的探讨脑血管狭窄患者Nf1基因的突变热点及突变方式。方法从全血中提取DNA,采用变性高效液相色谱分析(DHPLC)对5例脑血管狭窄病例进行Nf1全基因突变筛查;对DHPLC检测有差异洗脱峰型的区域进行测序分析。结果 5例无危险因素的脑血管狭窄患者Nf1全基因筛查发现:(1)1例出现5号外显子无义突变位点,c.541C-T;(2)5例均有14号和15号外显子之间内含子的基因突变;(3)4例有7号外显子c.702G-A的同义突变位点;(4)1例出现32号(c.4177G-A,Val-Ile)和46号外显子(c.6918T-G,Asn-Lys)的错义突变。结论在目前无其他基础疾病的脑血管狭窄患者体内的Nf1基因并非以稳定野生型存在,而是具有不同类型的突变,这些突变集中在第5、7、14、32和46等外显子上。Objective The results of a Chinese family affected by NF1 combined with cerebral vessel stenosis had revealed a nonsense mutation, c.541 C-T, in the NF1 gene（NM_001042492.2）. We aimed to screen the mutations for the full-length Nf1 gene and make a preliminary exploration on mutation hotspots and types of Nf1 gene in 5 cerebrovascular stenosis patients without other underlying disorders and causative factors. Methods ⅡDNA was extracted from whole blood. ⅡDenaturing high performance liquid chromatography（DHPLC）, a high throughput, non-hazardous and largely automated heteroduplex-based technique, was optimized for the rapid screening of the 60 exons and splice junctions of the Nf1 gene. ⅡMutable points were analyzed using sequencing analysis. Results After Nf1 gene screening for 5 cerebral vessels stenotic patients without underlying disorders and causative factors, results showed that：ⅡA nonsense mutation,c.541 C-T in exon 5 was identified for one patient; ⅡA base substitution, T/C or C/C in the intron between exon 14 and 15 was identified for all the cerebrovascular disease patients. The sense of this mutation was not clear. ⅡA same sense mutation, c.702 G-A in exon 7 was identified for four patients; ⅡTwo missense mutations, c.4177 G-A C（Val-Ile） in exon 32 and. c.6918 T-G（Asn-Lys） in exon 46 were identified for one patient. Conclusion Nf1 gene is not a stable wild type in patients with cerebrovascular stenosis but without other underlying disorders and causative factors. The gene can occur in different types of mutations. These mutations are concentrated in the 5 th, 7 th, 14 th, 32 nd and 46 th exon.

关 键 词：脑血管狭窄 NF1基因 DHPLC 突变分析 

分 类 号：R743[医药卫生—神经病学与精神病学]