采用二代测序对α/β复合型地中海贫血夫妇进行胚胎植入前遗传学诊断  

Preimplantation Genetic Diagnosis of α/β Complex Thalassemia by Next Generation Sequencing

作　　者：何天文[1] 卢建[1] 陈创奇[2] 周伟宁[1] 李静姝 董云巧[2] 杜丽[1] 尹爱华[1] HE Tian-Wen;LU Jian;CHEN Chuang-Qi;ZHOU Wei-Ning;LI Jing-Shu;DONG Yun-Qiao;DU Li;YIN Ai-Hua(Medical Genetics Center,Key Laboratory of Metabolic and Genetic Disease in Women and Children,Guangdong Women and Children Hospital,Guangzhou 511442,Guangdong Province,China;Reproductive Center of Guangdong Women and Children Hospital,Guangzhou 511442,Guangdong Province,China)

机构地区：[1]广东省妇幼保健院医学遗传中心,广东省妇幼代谢与遗传病重点实验室,广东广州511442 [2]广东省妇幼保健院生殖中心,广东广州511442

出　　处：《中国实验血液学杂志》2021年第4期1275-1279,共5页Journal of Experimental Hematology

基　　金：国家重点研发计划项目(2016YFC1000703)。

摘　　要：目的:探讨二代测序在α/β复合型地中海贫血夫妇胚胎植入前遗传学诊断的应用价值。方法:选择α-珠蛋白基因(HBA1、HBA2)和β-珠蛋白基因(HBB)编码区为目标区域,在基因上下游2M区域内选择高密度紧密连锁的单核苷酸多态位点作为遗传连锁标记,二代测序后选择有效单核苷酸多态位点构建夫妇单体型,确定夫妇携带致病基因的风险染色体。采用二代测序技术对胚胎HBA1、HBA2和HBB直接测序和构建单体型连锁分析进行植入前遗传学诊断。结果:胚胎HBA1、HBA2和HBB直接测序和构建单体型连锁分析结果显示,活检的6个囊胚中2个为α/β复合型地中海贫血,1个为β地中海贫血杂合携带,2个为α地中海贫血杂合携带,1个为中间型α地中海贫血。选择发育良好且植入前遗传学诊断结果提示可以移植的胚胎植入母体子宫后,足月分娩一健康婴儿。结论:采用二代测序技术可对α/β复合型地中海贫血夫妇进行胚胎植入前遗传学诊断,同时可以避免选择非整倍体胚胎而导致的流产问题。Objective: To explore the application value of next generation sequencing(NGS) in preimplantation genetic diagnosis of α/β complex thalassemia couple. Methods: The coding regions of α-globin genes(HBA1, HBA2)and β-globin gene(HBB) were selected as the target regions. The high-density and closely linked single nucleotide polymorphism(SNP) sites were selected as the genetic linkage markers in the upstream and downstream 2 M regions of the gene. After NGS, the effective SNP sites were selected to construct the haplotype of the couple, and the risk chromosome of the mutation carried by the couple was determined. The NGS technology was used to sequence the variations of HBA1, HBA2 and HBB directly and construct haplotype linkage analysis for preimplantation genetic diagnosis. Results: Direct sequencing and haplotype linkage analysis of HBA1, HBA2 and HBB showed that two of the six blastocysts were α/β complex thalassemia, one was β-thalassemia heterozygote, two were α-thalassemias heterozygotes, and one was intermediate α-thalassemia. A well-developed embryo underwent preimplantation genetic diagnosis was implanted into the mother’s uterus, and a healthy infant was born at term. Conclusion: Preimplantation genetic diagnosis can be carried out by NGS technology in α/β complex thalassemia couples, and abortion caused by aneuploid embryo selection can be avoided.

关 键 词：二代测序 α/β复合型地中海贫血 植入前遗传学诊断 

分 类 号：R556.61[医药卫生—血液循环系统疾病]