应用下一代测序技术对α地中海贫血进行胚胎植入前遗传学检测  

Successful preimplantation genetic detection for alpha thalassaemia by using next generation sequencing technology

作　　者：谢美娟 邓权衡 邓红辉 卢绍月 杨学习[2] 马强[2] 

机构地区：[1]广州市达瑞生物技术股份有限公司,广东广州510665 [2]南方医科大学检验与生物技术学院,广东广州510515

出　　处：《分子诊断与治疗杂志》2016年第6期367-374,共8页Journal of Molecular Diagnostics and Therapy

基　　金：广州市科技计划项目健康医疗协同创新重大专项(201400000004-4);广州市重大科技攻关项目子课题(2014Y2-00220);广州市科技计划产学研协同创新重大专项(201604020104);广东省科技计划项目公益研究与能力建设专项(2015A030401040)

摘　　要：目的探讨下一代测序(next generation sequencing,NGS)技术在α地中海贫血胚胎植入前遗传学检测中的应用。方法选取2对α地中海贫血--SEA缺失型携带者夫妇体外受精胚胎活检后的6个胚胎样本应用全基因组扩增(whole genome amplification,WGA)技术、下一代测序技术进行胚胎植入前遗传学检测,同时采用跨越断裂点荧光PCR(gap-PCR)进行平行对照检测。结果 2个家系6个胚胎样本的胚胎植入前遗传学诊断(preimplantation genetic diagnosis,PGD)结果分别为--SEA/αα母源携带、--SEA/--SEA、--SEA/αα母源携带、--SEA/--SEA、--SEA/αα母源携带和--SEA/--SEA;胚胎植入前遗传学筛查(preimplantation genetic screening,PGS)结果分别为45,XX,-5、46,XX、46,XY、47,XY,+1、46,XX和46,XY,+1,-2;Family 1 gap-PCR检测结果为父母均为SEA杂合子;E1为正常、E2为SEA纯合子、E3为正常;Family 2检测结果分别为:父母均为SEA杂合子;E4为SEA纯合子、E5为正常、E6为SEA纯合子。结论结果显示利用NGS不仅可以检测出23对染色体的核型,同时解决了单细胞扩增等位基因脱扣(allele drop-out,ADO)造成的假阳性和假阴性的风险,更具有市场潜力及应用前景。Objective To explore the application of next generation sequencing（NGS） for alphathalassemia in preimplantation genetic detection. Methods 2 couples with alpha-thalassemia Southeast Asia deletion（--SEA deletion） and their embryos cells obtained after embryonic biopsy were selected to test by whole genome amplification（WGA） and NGS. Gap-PCR was done to all samples as a series of parallel tests.Results The PGD results were--SEA /αα,--SEA/--SEA,--SEA/αα,--SEA/--SEA,--SEA /αα and--SEA /--SEA, respectively. And embryos with the abnormal α gene were confirmed from the maternal side according to genealogical analysis. The PGS results of 6 embryos from 2 families were 45, XX,-5; 46, XX; 46, XY; 47, XY,＋1; 46, XX and 46, XY＋1-2. Both husbands and wives of the 2 families gap-PCR results were heterozygotes with--SEA deletion. The results of gap-PCR were normal in embryos 1, 3 and 5 while--SEA deletion were found in embryos 2, 4 and 6 accordingly. Conclusion NGS not only can detect karyotype of 46 chromosomes but also avoid the risk of false positive and false negative caused by the allele drop-out（ADO）during the single cell amplification with a huge potential market and wide application prospect.

关 键 词：下一代测序 Α地中海贫血 胚胎植入前遗传学诊断 跨越断裂点荧光PCR 

分 类 号：R714.8[医药卫生—妇产科学]