常染色体隐性遗传视网膜色素变性CNGA1基因突变检测  

Detection of CNGA1 mutation in Chinese families with autosomal recessive retinitis pigmentosa

作　　者：熊世红[1] 赵堪兴[2] 王立[2] 陈薇樱[2] 王犁明[2] 王擎 

机构地区：[1]首都医科大学附属北京友谊医院 [2]天津市眼科医院 [3]Center for Molecular Genetics,Clevel and Clinic Foundation,Ohio44195,USA

出　　处：《眼科》2005年第4期261-264,共4页Ophthalmology in China

基　　金：国家自然科学基金杰出青年科学基金(39825510)

摘　　要：目的检测常染色体隐性遗传视网膜色素变性患者杆体αcGMP门离子通道基因(αcGMPgatedcationchannel,CNGA1)基因突变。设计对照性实验研究。研究对象35名常染色体隐性遗传视网膜色素变性(autosomalrecessiveretinitispigmentosa,ARRP)家系的先证者和55名散发病例。随机收集100名正常人作为对照。方法采集患者外周血,应用DNA分离试剂盒提取DNA,应用11对CNGA1基因引物进行聚合酶链反应(polymerasechainreaction,PCR),扩增CNGA1基因的全部编码区及内含子外显子的拼接区。利用单链构象多态性(singlestrandconformationpolymorphism,SSCP)技术,将PCR产物进行10%非变性聚丙烯酰胺凝胶电泳,用硝酸银染色,观察有无变异带。如果发现变异带,再将该PCR产物进行DNA测序。主要指标通过PCRSSCP和DNA测序技术,发现CNGA1基因突变。结果未发现CNGA1基因突变。结论CNGA1基因突变在国人RP患者中的致病情况有待进一步研究。Objective To detect α-cGMP-gated cation channel （CNGA 1） mutation in Chinese families with autosomal recessive retinitis pigmentosa （ARRP）. Design Comparative experimental study. Participants 35 ARRP probands and 55 sporadic RP patients were analyzed, 100 non-related normal subjects were collected as controls. Methods Genetic DNA was prepared from peripheral blood leukocytes using DNA Isolation Kits for Mammalian Blood. The complete coding region and exon-intron splice sites of CNGA 1 gene were amplified with polymerase chain reaction （PCR）. DNA single-strand conformation polymorphism （SSCP） technique was used to screen CNGA 1 gene mutation. PCR products were loaded on a 10% polyacryarnide gel. After electrophoresis the polyacryamid gel was silver stained. When a variant band was discovered with SSCP electrophoresis, the variant hand were analyzed with sequencing PCR-amplified DNA. Main Outcome Measures Identifying mutations in the CNGA 1 gene using PCR-SSCP and DNA sequencing techniques. Results No mutation was found in these patients. Conclusions Further study should he required for knowing CNGA 1 gene mutation in Chinese patients with RP.

关 键 词：视网膜色素变性 CNGA1基因 基因突变 

分 类 号：R774.1[医药卫生—眼科]