CAJ | 학술논문

目的探讨利用目标外显子捕获结合高通量测序技术检测胎儿骨骼畸形致病基因的效果.方法 2009年7月至2014年7月在解放军总医院进行产前检查的孕妇均于妊娠18～24周和/或30～32周行常规胎儿超声检查,其中10例孕妇超声检查提示胎儿骨骼畸形,纳入研究.超声引导下行羊膜腔穿刺术或脐静脉穿刺术留取羊水或脐带血,行染色体核型分析.取羊水或脐带血,或引产后取胎儿组织,或分娩后留取新生儿血样,同时留取胎儿/新生儿父母的静脉血,提取DNA,利用目标外显子捕获结合高通量测序技术进行248种骨骼畸形相关基因的筛查,对可疑位点进行一代测序验证.结果 10例骨骼畸形胎儿染色体核型分析均未见异常.目标外显子捕获结合高通量测序技术表明其中3例胎儿存在Ⅰ型胶原α1链(collagen,type Ⅰ,alpha-l,COL1A1)基因出现杂合突变,分别为c.3307G＞A、c.1706G＞C及c.2101G＞A.胎儿父母双方均进行了基因组DNA一代测序,未发现突变.3例胎儿检出成纤维细胞生长因子受体3(fibroblast growth factor 3,FGFR3)基因杂合突变,其中2例为c.1138G＞A,另一例为c.1118A＞G.1例胎儿检出埃利伟综合征(Ellis-van Creveld syndrome,EVC)基因c.884C＞G和c.982C＞T复合杂合突变,一代测序证实2个突变位点分别来自于胎儿父母.另外3例胎儿未发现明确致病基因. 结论利用目标外显子捕获结合高通量测序技术可为部分骨骼异常胎儿明确致病基因.
목적 탐토이용목표외현자포획결합고통량측서기술검측태인골격기형치병기인적효과.방법 2009년7월지2014년7월재해방군총의원진행산전검사적잉부균우임신18～24주화/혹30～32주행상규태인초성검사,기중10례잉부초성검사제시태인골격기형,납입연구.초성인도하행양막강천자술혹제정맥천자술류취양수혹제대혈,행염색체핵형분석.취양수혹제대혈,혹인산후취태인조직,혹분면후류취신생인혈양,동시류취태인/신생인부모적정맥혈,제취DNA,이용목표외현자포획결합고통량측서기술진행248충골격기형상관기인적사사,대가의위점진행일대측서험증.결과 10례골격기형태인염색체핵형분석균미견이상.목표외현자포획결합고통량측서기술표명기중3례태인존재Ⅰ형효원α1련(collagen,type Ⅰ,alpha-l,COL1A1)기인출현잡합돌변,분별위c.3307G＞A、c.1706G＞C급c.2101G＞A.태인부모쌍방균진행료기인조DNA일대측서,미발현돌변.3례태인검출성섬유세포생장인자수체3(fibroblast growth factor 3,FGFR3)기인잡합돌변,기중2례위c.1138G＞A,령일례위c.1118A＞G.1례태인검출애리위종합정(Ellis-van Creveld syndrome,EVC)기인c.884C＞G화c.982C＞T복합잡합돌변,일대측서증실2개돌변위점분별래자우태인부모.령외3례태인미발현명학치병기인. 결론 이용목표외현자포획결합고통량측서기술가위부분골격이상태인명학치병기인.
Objective To investigate the feasibility of targeted exome capture with high throughput sequencing in gene mutations related to fetal skeletal dysplasia.Methods From July 2009 to July 2014,ten fetuses with skeletal dysplasia were identified by ultrasound screening at 18-24 and/or 30-32 gestational weeks in the Chinese PLA General Hospital.Amniotic fluid or cord blood was collected for karyotyping.Amniotic fluid or cord blood,fetal tissues after labor induction,and blood samples from neonates and parents were collected and analyzed for 248 genes associated with fetal skeletal dysplasia using targeted exome capture with high throughput sequencing.Detected gene mutations were confirmed by direct Sanger sequencing reactions.Results No abnormal karyotypes were found in the ten fetuses.Three fetuses carried collagen,type Ⅰ,alpha-1 (COLIA1) gene mutations of c.3307G＞A,c.1706G＞C and c.2101G＞A,respectively.No mutations were found in their parents,which were confirmed by direct Sanger sequencing reactions.Another three fetuses carried fibroblast growth factor 3 (FGFR3) gene mutations of c.1138G＞A and c.1118A＞G,respectively.One fetus carried compound heterozygous Ellis-van Creveld syndrome (EVC) gene mutations of c.884C＞G and c.982C＞T from her parents and were confirmed by direct Sanger sequencing reactions.No causative mutations were found in the remaining three cases.Conclusion Targeted exome capture with high throughput sequencing is a new approach for identifying causative gene mutations in fetal skeletal dysplasia.