摘要
目的探讨二代测序在Marfan综合征家系的植入前遗传学诊断应用价值和优势。方法针对2016年10月在广东省妇幼保健院医学遗传中心就诊的1例Marfan综合征家系行外显子捕获测序筛查微纤维蛋白(FBN1)基因突变位点,筛查结果通过Sanger测序进行验证,明确基因突变位点。选择FBN1基因编码区及外显子-内含子交界为目标区域,在该基因上下游2 M区域内选择100个高密度紧密连锁的单核苷酸多态位点(SNP)作为遗传连锁标记,构建夫妇单倍型。采用二代测序对胚胎的突变位点直接测序和单体型连锁分析进行植入前遗传学诊断。结果男方存在FBN1基因c.247+1G>A的致病性突变,经直接测序和单体型连锁分析,患者夫妇的5个囊胚中3个正常,2个致病。选择发育良好且遗传学检测正常的胚胎植入母体子宫,足月分娩一健康婴儿。结论采用二代测序对Marfan综合征家系进行植入前遗传学诊断可以阻断此单基因病在该家系中的再发风险,还可以避免选择非整倍体胚胎而导致的流产问题,是Marfan综合征出生缺陷的有效预防手段。
Objective To explore the value and advantage of next-generation sequencing in preimplantation genetic diagnosis of Marfan syndrome.Methods A Marfan syndrome family was screened for the FBN1 gene mutation site by exon capture sequencing in the medical genetics center of Guangdong Women and Children Hospital in October 2016.The screening results were verified by Sanger sequencing,and the mutation site was identified.The coding region and exon intron junction of the FBN1 gene were selected as the target region,and 100 high-density closely linked single nucleotide polymorphisms(SNP)were selected as the genetic linkage markers in the upstream and downstream 2 m regions of the FBN1 gene to construct the couple haplotypes.Direct sequencing of mutation sites and haplotype linkage analysis by next generation sequencing were used for preimplantation genetic diagnosis.Results There was a pathogenic mutation of the FBN1 gene c.247+1 g>A in the male side.By direct sequencing and haplotype linkage analysis,3 of the 5 blastocysts of thepatients were normal and 2 were pathogenic.A well-developed and genetically normal embryo was implanted into the mother.s uterus,and a healthy baby was born at full term.Conclusion preimplantation genetic diagnosis of Marfan syndrome family by next generation sequencing can block the risk of recurrence of this single gene disease in the family,and can also avoid the abortion caused by the selection of aneuploid embryos,which is an effective prevention method for birth defects of Marfan syndrome.
作者
何天文
卢建
陈创奇
刘顿
丁红珂
董云巧
杜丽
尹爱华
HE Tianwen;LU Jian;CHEN Chuangqi;LIU Dun;DING Hongke;DONG Yunqiao;DU Li;YIN Aihua(Medical Genetics Center of Guangdong Women and Children Hospital,Guangzhou,Guangdong,China,511442;Maternal and Children Metabolic-Genetic Key Laboratory of Guangdong,Guangzhou,Guangdong,China,511442;Reproductive center of Guangdong Women and Children Hospital,Guangzhou,Guangdong,China,511442)
出处
《分子诊断与治疗杂志》
2020年第8期995-1000,共6页
Journal of Molecular Diagnostics and Therapy
基金
国家重点研发计划项目(2016YFC1000703)。
