【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。更多还原

【Abstract】 Objective To detect α-cGMP-gated cation channel (CNGA1) 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 CNGA1 gene were amplified with polymerase chain reaction (PCR).DNA single-strand conformation polymorphism(SSCP) technique was used to screen CNGA1 gene mutation.PCR products were loaded on a 10% polyacryamide gel.After electrophoresis the polyacryamid gel was silver stained.When a variant band was discovered with SSCP electrophoresis,the variant band were analyzed with sequencing PCR-amplified DNA.Main Outcome Measures Identifying mutations in the CNGA1 gene using PCR-SSCP and DNA sequencing techniques.Results No mutation was found in these patients.Conclusions Further study should be required for knowing CNGA1 gene mutation in Chinese patients with RP.更多还原