CAJ | 학술논문

背景先天性白内障是造成儿童盲和弱视的重要原因,其中约50％的先天性白内障具有遗传性.目的应用眼遗传病外显子结合目标区域捕获测序芯片检测一常染色体显性遗传先天性白内障家系的致病基因.方法于2011年在宁夏眼科医院收集一回族常染色体显性遗传先天性白内障家系,采集家系中患者及表型正常成员的临床资料.对家系成员进行眼科检查,抽取患者、表型正常家系成员及300名正常对照者的外周静脉血各5 ml,提取DNA,利用眼遗传病外显子结合目标区域捕获测序芯片筛查和检测候选致病突变位点.采用PCR和直接测序法对家系成员和正常对照者进行突变位点验证,最终确定致病突变位点.结果该家系共6代61名成员,均为回族,先天性白内障患者18例,为5代遗传,符合常染色体显性遗传特征.患者中合并眼球震颤和斜视者7例,合并高度近视者4例,来诊前均已实施白内障摘出术.利用眼遗传病外显子结合目标区域捕获测序芯片检测结合生物信息学方法筛查后共得到8个候选致病突变位点,其中5个在非编码区,3个在编码区,通过PCR和直接测序法验证确定CRYGD基因上的P24T突变是该家系的致病突变位点.该突变与家系内所有患者表型共分离,在家系表型正常者及300名正常对照者均未发现此突变.结论外显子结合目标区域捕获测序技术快速检测CRYGD基因P24T突变为该先天性白内障家系致病突变,该技术为临床表型多样、致病基因众多的先天性白内障的致病基因检测提供新的手段.
배경 선천성백내장시조성인동맹화약시적중요원인,기중약50％적선천성백내장구유유전성.목적 응용안유전병외현자결합목표구역포획측서심편검측일상염색체현성유전선천성백내장가계적치병기인.방법 우2011년재저하안과의원수집일회족상염색체현성유전선천성백내장가계,채집가계중환자급표형정상성원적림상자료.대가계성원진행안과검사,추취환자、표형정상가계성원급300명정상대조자적외주정맥혈각5 ml,제취DNA,이용안유전병외현자결합목표구역포획측서심편사사화검측후선치병돌변위점.채용PCR화직접측서법대가계성원화정상대조자진행돌변위점험증,최종학정치병돌변위점.결과 해가계공6대61명성원,균위회족,선천성백내장환자18례,위5대유전,부합상염색체현성유전특정.환자중합병안구진전화사시자7례,합병고도근시자4례,래진전균이실시백내장적출술.이용안유전병외현자결합목표구역포획측서심편검측결합생물신식학방법사사후공득도8개후선치병돌변위점,기중5개재비편마구,3개재편마구,통과PCR화직접측서법험증학정CRYGD기인상적P24T돌변시해가계적치병돌변위점.해돌변여가계내소유환자표형공분리,재가계표형정상자급300명정상대조자균미발현차돌변.결론 외현자결합목표구역포획측서기술쾌속검측CRYGD기인P24T돌변위해선천성백내장가계치병돌변,해기술위림상표형다양、치병기인음다적선천성백내장적치병기인검측제공신적수단.
Background Congenital cataract is an important cause of blindness and amblyopia in children,and about 50％ of congenital cataract is hereditary.Objective The aim of this study was to determine the diseasecausing gene of one Hui congenital cataract pedigree by using exon combined target region capture sequencing chip of eye diseases.Methods This study was approved by Ethic Committee of Ningxia People's Hospital and followed Declaration of Helsinki.One Hui congenital cataract pedigree was recruited in Ningxia Eye Hospital in 2011.All the disease history of the members in this family were collected and recorded,and the eye examinations were performed.The peripheral blood specimens were collected from family members and 300 healthy individuals for the extraction of DNA.Exon combined target region capture sequencing chip of eye diseases was used to screen the candidate diseasecausing mutations,then PCR and direct sequencing were used to confirm the disease-causing mutations.Results This H ui family included 61 members of 6 generations,and 18 patients were diagnosed in serial 5 passages,conforming to autosomal dominant inheritance pattern.Among 18 cataract patients,7 individuals were associated with nystagmus and strabismus,and 4 patients had high myopia.Eight candidate pathogenetic mutations were detected by exon combined target region capture sequencing chip of eye diseases and bioinformatics method,with 5 mutations in noncoding regions and 3 in coding regions.The mutation P24T of CYRGD gene was confirmed as pathogenic mutation of this pedigree by using PCR and direct sequencing methods.These mutations co-segregated with affected members of the family,and the mutations were not found in the unaffected family members and 300 unrelated controls.Conclusions P24T of CYRGD gene mutation is confirmed as pathogenic mutation of this pedigree.Exon combined target region capture sequencing chip provides a new approach to detect disease-causing mutations of congenital cataract with diversity clinical phenotypes.