应用变性高效液相色谱技术快速诊断儿童型脊髓性肌萎缩症(英文)  

Rapid diagnosis of spinal muscular atrophy using denaturing high-performance liquid chromatography

作　　者：陈万金[1] 吴志英[1] 王柠[1] 林珉婷 慕容慎行[1] 

机构地区：[1]福建医科大学附属第一医院神经内科,福州350005 [2]福建省神经病学研究所

出　　处：《中华医学遗传学杂志》2005年第3期291-293,共3页Chinese Journal of Medical Genetics

基　　金：福建省重大项目基金(2002Y001)~~

摘　　要：目的介绍变性高效液相色谱(denaturinghighperformanceliquidchromatography,DHPLC)技术在儿童型脊髓性肌萎缩症(spinalmuscularatrophy,SMA)基因诊断中的应用。方法PCR扩增25名正常人、1份标准样品及25例SMA患者运动神经元生存基因(survivalmotorneuron,SMN)第7外显子及其侧翼区域,PCR产物变性、复性后直接上样于DHPLC系统。通过改变A、B缓冲液的比例来分离各种DNA成份。结果各种不同DNA成份以色谱峰的形式表现出来。23名正常标本呈现3个峰,依次为SMN1/SMN2异源双链峰、SMN2同源双链峰、SMN1同源双链峰。2名正常标本及1份标准品只有SMN1峰,表明缺失了SMN2。22例SMA患者只有SMN2峰,表明缺失了SMN1。另3例SMA患者呈现3个峰,表明无SMN1或SMN2缺失。结论DHPLC诊断SMA具有敏感、准确、快速、简便等优点。Objective To introduce the application of de n aturing high-performance liquid chromatography(DHPLC) in the diagnosis of child hood type spinal muscular atrophy(SMA). Methods Exon 7 and flan king area of survival motor neuron (SMN) gene were amplified by PCR in 1 s tandard sample,25 normal individuals and 25 patients with SMA. The PCR products were then directly loaded onto the DHPLC system after denaturing and annealing. Different DNA segments were separated by changing the concentration of buffer A relative to that of buffer B. Results Different DNA segments w ere separable on the DHPLC chromatogram. Three peaks including SMN1/SMN2 heteroduplex peak, SMN2 homoduplex peak and SMN1 h o moduplex peak were detected in 23 out of 25 normal individuals. Only SMN1 homoduplex peak was detected in 2 normal individuals and the standard samp le, indicating the deletion of SMN2. On the contrary, only the SMN2 homoduplex peak was detected in 22 out of 25 patients with SMA, indi cating deletion of SMN1. The three peaks as those of normal individ u als were detected in the other 3 patients, indicating no SMN1 or SMN2 deletion. Conclusion As a new technology for diag nosing SMA, DHPLC is sensitive, accurate, rapid and convenient.

关 键 词：变性高效液相色谱技术 快速诊断 儿童 脊髓性肌萎缩症 运动神经元 

分 类 号：R746[医药卫生—神经病学与精神病学]