单精子SNP单体型在1例Ⅱ型神经纤维瘤病患者胚胎植入前遗传学检测中的应用  

Base on Single Sperm SNP Haplotype in the Preimplantation Genetic Examination of A Patient with Neurofibromatosis TypeⅡ

作　　者：周永红 谢雨莉 付志红 韩婵琳 焦淑静 李雪梅 邓天勤 ZHOU Yong-hong;XIE Yu-li;FU zhi-hong;HAN chan-lin;JIAO Shu-jing;LI Xue-mei;DENG Tian-qin(Center of Reproductive Medicine,Shenzhen Maternity and Child Healthcare Hospital of South Medical University,Shenzhen 518028,Guangdong Province,China;Center for Neonatal Disease Screening,Shenzhen Maternity and Child Healthcare Hospital of South Medical University,Shenzhen518028,Guangdong Province,China;Yikon Genomics Company,Suzhou215021,Jiangsu Province,China)

机构地区：[1]南方医科大学附属深圳妇幼保健院生殖医学中心,广东深圳518028 [2]南方医科大学附属深圳妇幼保健院新生儿疾病筛查中心,广东深圳518028 [3]苏州亿康医学检验有限公司,江苏苏州215021

出　　处：《罕少疾病杂志》2023年第12期1-3,共3页Journal of Rare and Uncommon Diseases

基　　金：深圳市医学重点学科建设经费资助(SZXK031);深圳市科创委基础研究面上项目(JCYJ20220530155011024)。

摘　　要：目的探讨单精子SNP单体型在新发突变的胚胎植入前遗传学检测中意义。方法对一名由NF2基因c.861delC杂合突变的常染色体显性的神经纤维瘤Ⅱ型的患者,运用高通量测序技术建立单体型。用机械法分离单精子并行全基因组扩增,采用ASA基因芯片,分析致病基因上游以及下游1M内的单核苷酸多态性(SNP)位点,确定染色体单体型是否携带突变。活检五份囊胚滋养层细胞样本,采用全基因组扩增及高通量测序,鉴定胚胎携带或不携带致病突变,选择未携带致病基因和染色体正常的囊胚进行移植,在孕中期(18周)行产前诊断,确认胎儿染色体核型、结构及是否携带致病突变。结果通过ASA基因芯片检测,共筛选出30个SNP位点,成功建立SNP单体型。检测显示一枚胚胎携带致病突变,四枚未携带,经孕中期产前诊断,胎儿未携带NF2基因c.861delC杂合突变。结论对缺乏先证者,携带新发突变男性患者,可通过ASA基因芯片构建单精子SNP单体型的检测策略,进行胚胎植入前遗传学检测,阻断致病基因的传递。Objective To investigate the value of single sperm SNP haplotype in preimplantation genetic testing.Methods In a patient with autosomal dominant neurofibromatosis type II by a heterozygous mutation of c.861delC in the NF2 gene,haplotypes were established using high-throughput sequencing.Sperm were isolated and genome-wide amplified by mechanical isolation,and single nucleotide polymorphism(SNP)sites upstream as well as within 1 M downstream of the gene were analyzed using ASA GeneChips to determine chromosomal haplotypes carrying and not carrying the mutation.Five blastocyst trophoblast cell samples were biopsied,and whole genome amplification and high-throughput sequencing were used to identify whether the embryos carried the causative mutation or not,to select suitable blastocysts for transfer,and to perform prenatal diagnosis at 18 weeks of gestation to confirm whether the fetus carried the causative mutation or not.Results A total of 30 SNP were selected by single sperm SNP detection,and haplotype was successfully constructed.Haplotype analysis before implantation suggested that one embryo carried pathogenic mutation,and four did not.Amniotic fluid test in the second trimester confirmed that the fetus did not carry heterozygous mutation NF2 gene C.861delc.Conclusion For male with de novo pathogenic variants,single sperm SNP haplotype linkage analysis,preimplantation genetic testing,and normal embryos for transfer can be performed to block the vertical transmission of pathogenic genes.

关 键 词：神经纤维瘤病2型 胚胎植入前遗传学检测 精子 单体型 

分 类 号：R394[医药卫生—医学遗传学]