一种简化的检测SMN1基因纯合性缺失的方法  

A Simplified Approach for Detecting Homologous Deletion of SMN1 Genes in Spinal Muacular Atrophy

作　　者：李晓侨[1] 姚凤霞[2] 苏亮[1] 韩娟娟[2] 孟岩[1] 王铮[1] 彭园园[1] 佃艳[1] 周青[1] 黄尚志[1,2] 

机构地区：[1]中国医学科学院基础医学研究所医学遗传学系/北京协和医学院基础学院WHO遗传病社区控制合作中心,北京100005 [2]北京协和医院中心实验室,100730

出　　处：《医学研究杂志》2009年第5期29-32,137,共5页Journal of Medical Research

基　　金：科技部"十一五"国家科技支撑计划项目(2006BIA05A08);北京市科学技术委员会研发攻关类基金资助项目(D0906005040491)

摘　　要：目的建立一种更加准确、快捷的方法,简便地诊断纯合缺失型脊髓性肌萎缩症。方法应用双侧等位基因特异聚合酶链反应(AS-PCR)进行SMN1基因的特异扩增,并用另1个无关基因作内对照,通过聚丙烯酰胺凝胶电泳(PAGE)分析,确定患儿是否为纯合缺失型脊髓性肌萎缩症。结果双重AS-PCR产物,通过PAGE或琼脂糖凝胶电泳,可准确判读患儿是否存在SMN1基因外显子7纯合性缺失。结论相对于以往常规使用的PCR-酶切或变性高效液相层析(DHPLC)方法,本研究所建立的方法不需要复杂的后续分析手段,能够更准确、快捷、简便地诊断纯合缺失型脊髓性肌萎缩症患儿。Objective To develop a rapid,reliable and convenient approach for diagnosing the homozygous deletion of SMN1 gene. Methods SMNI gene was amplified specifically with double allele - specific PCR（ AS - PCR）. Meanwhile, one inrelevant gene was amplified as internal control by PAGE and agarose gel electrophoresis analysis to determine whether the sick children were with homozygous deletion of SMN1 genes. Results The homozygous deletion of exon7 in SMN1 gene was identified by agarose gel electrophoresis or PAGE accurately. Conclusion Compared to PCR -RFLP and DHPLC used in the past, this approach can diagnose homozygous deletion of SMA much more accurate, easier and more convenient without completed following analyses.

关 键 词：脊髓性肌萎缩 等位基因特异聚合酶链反应 运动神经元存活基因 聚丙烯酰胺凝胶电泳 琼脂糖凝胶电泳 

分 类 号：R734.2[医药卫生—肿瘤] R382.31[医药卫生—临床医学]