眼病候选基因芯片在一汉族视网膜色素变性家系分子遗传学中的应用  

Next generation sequencing based molecular genetic analysis of a Chinese Han family with autosomal retinitis pigmentosa

作　　者：周晓敏[1] 黄辉 王瑛[1] 吴静 樊宁[3] 蒋善明 刘旭阳[3] 

机构地区：[1]四川大学华西医院医教部/眼科学研究室,成都610041 [2]深圳华大基因研究院,518000 [3]暨南大学附属深圳市眼科医院,518000

出　　处：《中华实验眼科杂志》2015年第8期699-703,共5页Chinese Journal Of Experimental Ophthalmology

摘　　要：背景 视网膜色素变性（RP）是一种累及视网膜光感受器细胞及色素上皮细胞的遗传性致盲眼病.RP的发病机制及临床特征较复杂,具有遗传异质性和临床异质性.随着基因组学的迅猛发展,越来越多的研究手段应用于RP致病基因筛查.目的 通过眼科基因芯片测序方法探讨一常染色体遗传RP家系临床表型及其基因突变情况.方法 于2013年6月在重庆市荣昌县收集一汉族RP家系,对该家系所有患者进行眼科检查确诊后,抽取12名家系成员外周静脉血各1 ml,应用华大基因眼科芯片目标区域捕获技术进行基因突变检测.该基因芯片覆盖了眼病相关的基因编码区（包括59个RP候选基因）,选择家系内2例RP患者（Ⅱ5、Ⅱ7）的DNA样本进行目标区域捕获测序.通过生物信息学技术对测序结果进行分析,对共有的变异位点进行Sanger测序验证.结果 该家系为常染色体遗传的RP家系.通过基因芯片分析发现该家系Ⅱ5和Ⅱ7患者存在2个共有基因突变：USH2A （c.3065T＞C,p.Phe1022Ser）突变和PDE6A（c.1699G＞A,p.Ala1319Gly）突变,家系其他成员检测结果表明2个基因突变未与疾病共分离.该眼科基因芯片高通量测序技术虽然未定位该家系致病基因,但快速排除了RP常见候选基因,为进一步分析奠定了研究基础.结论 采用基于目标区域捕获测序的眼科基因芯片技术可以快速、准确地筛查RP常见候选基因,是眼科疾病遗传研究的一项适用且高效的新方法.Background Retinitis pigmeutosa （RP） is a progressive inheritance disease.It is characterized by highly genetical and phenotypical heterogeneity.With the rapid development of genomics,new methods are applied to the genetic screening of RP.Objective This study was to characterize the clinical features of a Chinese family with autosomal RP and to screen the candidate genes.Methods Twelve members from this family were included in the study.All participants underwent complete ophthalmologic examinations.Targeted-capture next generation sequencing （NGS） based molecular genetic analysis was performed on two patients of this RP family（Ⅱ5,Ⅱ 7）.The DNA sample from the two patients was separately sequenced using custom capture gene chip,which includes 59 retinal disease genes.The sequencing results were analyzed by bioinformatics technology.Identified variations were verified in the rest family members by PCR and Sanger sequencing.This study was approved by Ethic Committee of West China Hospital,and informed consent was obtained from the subjects.Results Four members of this family were diagnosed as RP,and the rest were asymptomatic.Missense mutation （c.3065T〉C,p.Phe1022Ser） in USH2A and missense mutation （c.1699G〉A,p.Ala1319Gly） in PDE6A were found in two patients （Ⅱ 5 and Ⅱ7）.The variants were not co-segregated with the phenotype of this family.The causative mutation was not found by the targeted-capture NGS based eye disease chip,but it ruled out a large number of candidate genes for RP.Conclusions Our study suggests that targeted-capture NGS based eye disease chip can quickly detect mutations in known RP genes.It can be a new applicable and efficient method for molecular genetic analysis of ocular disease.

关 键 词：视网膜色素变性 基因 二代测序 分子遗传学 家系分析 

分 类 号：R774.1[医药卫生—眼科]