摘要
目的探讨外显子靶向捕获结合高通量测序技术在先天性白内障基因诊断中的应用价值方法收集1个先天性白内障家系先证者外周血,提取基因组DNA,并制备基因组文库。用GenCap基因序列捕获技术靶向富集文库中28个先天性白内障已知致病基因的外显子并进行高通量测序。通过生物信息学分析筛选致病突变。用Sanger测序验证相应致病基因突变位点。结果高通量测序获得了712.29 Mb的原始数据,测序数据经过生物信息学分析,发现CRYAA基因突变位点c.145C>T,突变导致CRYAA 49位精氨酸残基突变为半胱氨酸残基。Sanger测序验证了高通量测序的结果。结论利用靶向捕获结合高通量测序技术可鉴定先天性白内障的致病基因突变,对先天性白内障的临床基因诊断具有潜在的应用价值。
Objective To elucidate the usefulness of targeted exome sequencing for genetic diagnosis of congenital cataract(CC).Methods The whole blood from the proband of a Chinese family with CC was collected.Genomic DNA was extracted and the library was constructed.The coding exons and flanking regions of 28 genes related to CC were captured,followed by high-throughput sequencing.The sequencing data were analyzed by established bioinformatics pipeline.V alidation was performed by Sanger sequencing.Results 712.29 Mb raw data was obtained through TES.A heterozygous non-synonymous mutation c.145C>T in the CRYAA gene was identified after bioinformatics analysis.The mutation caused the substitution of an Arginine residue by a Cysteine residue at position 49 of CRYAA.The result was confirmed by Sanger sequencing.Conclusions Targeted exome sequencing is a rapid and cost-efficient method for the genetic diagnosis of CC.
作者
马明福
崔蓉
赵乐天
李练兵
程志
MA Mingfu;CUI Rong;ZHAO Letian;LI Lianbing;CHENG Zhi(Chongqing Population and Family Planning Science and Technology Research Institute,Key Laboratory of Birth Defects and Reproductive Health of National Health and Family Planning Commission,Chongqing 400020,China;Department of Cell Biology and Genetics,Chongqing Medical University,Chongqing 400016,China)
出处
《中国实用眼科杂志》
2018年第4期333-336,共4页
Chinese Journal of Practical Ophthalmology
基金
重庆市科委资助项目(2012CSTC-jbby-01704,CSTC,2009CA5001)。
