目标外显子捕获技术在胎儿骨骼畸形基因检测中的应用  

Application of targeted exome capture in identifying fetal skeletal malformation mutations

作　　者：张蔓丽[1] 卢彦平[1] 李芮冰 叶明侠[1] 黄柯[1] 游艳琴[1] 汪淑娟[1] 汪龙霞[3] 李亚里[1] 

机构地区：[1]解放军总医院妇产科,北京100853 [2]解放军总医院临检科,北京100853 [3]解放军总医院超声科,北京100853

出　　处：《中华围产医学杂志》2015年第5期334-338,共5页Chinese Journal of Perinatal Medicine

摘　　要：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.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.

关 键 词：骨疾病 发育性 肢畸形 先天性 外显子 高通量核苷酸序列分析 基因检测 超声检查 产前 

分 类 号：R714.5[医药卫生—妇产科学]