基于新一代测序的SNP单体型分析在先天性挛缩性蜘蛛样指症PGD中的应用  

Application of NGS-based SNP haplotyping for preimplantation genetic diagnosis of congenital contractural arachnodactyly

作　　者：陈林君[1] 刁振宇[1] 徐志鹏[1] 周建军[1] 颜桂军[1] 孙海翔[1] 

机构地区：[1]南京大学医学院附属鼓楼医院生殖医学科,南京210008

出　　处：《生殖医学杂志》2017年第1期59-63,共5页Journal of Reproductive Medicine

摘　　要：目的探讨基于新一代测序(NGS)的单核苷酸多态性(SNP)单体型分析在先天性挛缩性蜘蛛样指症(CCA)种植前遗传学诊断中的有效性。方法对活检的滋养层细胞采用多重置换扩增(MDA)的方法扩增全基因组,应用基于NGS的SNP单体型分析和直接测序两种方法对MDA产物进行CCA的种植前遗传学诊断(PGD)。结果应用MDA方法对活检的4枚囊胚的滋养层细胞成功地进行了全基因组扩增;通过基于NGS的SNP单体型分析发现,其中两枚是未感染CCA的囊胚,另两枚是感染CCA的囊胚;单体型分析结果与直接测序法结果一致。取卵5个月后患者移植一枚基因型正常的冷冻囊胚,于妊娠的第38周经剖宫产成功分娩一体重为2 850g的健康婴儿。结论基于NGS的SNP单体型分析是单基因病的种植前遗传学诊断的有效筛查工具。Objective： To investigate the effectiveness of next generation sequencing （NGS）-based single nucleotide polymorphism （SNP） haplotyping for preimplantation genetic diagnosis （PGD） of congenital contractural arachnodactyly （CCA）. Methods： Multiple displacement amplification （MDA） was performed for whole genome amplification （WGA） of biopsied trophectoderm cells. NGS-based SNP haplotyping and direct mutation detection by sequencing were used for PGD of CCA. Results： MDA was successfully performed for WGA of biopsied trophectoderm cells from four blastocysts. NGS-based SNP haplotype found that two blastocysts were unaffected by CCA, while another two blastocysts were affected by CCA. These diagnostic results were consistent with the results of direct mutation detection by sequencing. One blastocyst was transferred in the subsequent frozen embryo transfer （FET） cycle five months after ooeyte retrieved. A weight of 2 850 g of healthy baby was born by cesarean section at the 38^th week of gestation. Conclusions： NGS-based SNP haplotyping is an effective screening tool for PGD of monogenic disorders.

关 键 词：种植前遗传学诊断 先天性挛缩性蜘蛛样指症 多重置换扩增 新一代测序 单核苷酸多态性 

分 类 号：R[医药卫生]