赣南地区α-地贫和β-地贫植入前胚胎遗传学检测技术平台建立及临床应用  

Establishment and clinical application of preimplantation genetic testing platform forα-thalassemia andβ-thalassemia in southern Jiangxi area

作　　者：岳林林 何颖 黄云 帅妹 刘小舟 YUE Linlin;HE Ying;HUANG Yun;SHUAI Mei;LIU Xiaozhou(Ganzhou Maternity and Child Health Care Hospital,Ganzhou,Jiangxi Province,341000)

机构地区：[1]江西省赣州市妇幼保健院,341000

出　　处：《中国计划生育学杂志》2022年第11期2526-2530,2536,共6页Chinese Journal of Family Planning

基　　金：江西省卫生健康委科技计划(20194063)。

摘　　要：目的:运用MALBAC单细胞测序技术结合SNP连锁分析,建立赣南地区α-地中海贫血(简称α地贫)和β-地中海贫血(简称β地贫)植入前胚胎遗传学检测(PGT)技术平台并进行临床应用观察。方法:对12个α-地贫和8个β-地贫患者家系进行分析,构建家系单体型。进行了9个α-地贫和6个β-地贫的PGT治疗周期。依据胚胎拷贝数变异和致病位点携带情况选用可移植胚胎,在妊娠中期抽羊水做产前诊断验证。用废弃的检测出拷贝数变异或携带双方致病位点的不可移植胚胎进行室内和室间质控。结果:本实验室胚胎CNV/SNP建库测序结果与第三方实验室检测结果报告一致;通过两种单细胞全基因组扩增方法的结果比对及分析,显示MDA单细胞全基因组扩增方法的CNV/SNP建库失败率较高。结论:选择较为优势的MALBAC方法成功建立了赣南地区的地贫的胚胎植入前检测平台。Objective:To establish the preimplantation genetic testing(PGT)platform forα-thalassemia andβ-thalassemia by implanting MALBAC-NGS technique combining with SNP serial analysis in southern Jiangxi area,and to observe its clinical application.Methods:12 families withα-thalassemia and 8 families withβ-thalassemia were analyzed to build their pedigree haplotypes.Totally 9 PGT treatment periods of the patients withα-thalassemia and 6 PGT treatment periods of the patients withβ-thalassemia were completed.The transferable embryos were selected according to the copy number variation and the carrying status of pathogenic loci of chromosome of the embryos,and the amniotic fluid sampling of the pregnant women during the second trimester of pregnancy were obtained for prenatal diagnosis and verification.Internal quality control and external quality control were performed based on the embryos which couldn’t be qualified transplantation due to the embryos with both the copy number variation and the carrying status of pathogenic loci of chromosome of their parents.Results:The results of the established database of the embryo CNV-sequencing/SNP-sequencing techniques in this laboratory were consistent with those reported by the third-party laboratories.The comparison and analysis of the results of two single-cell genome-wide amplification methods showed that MDA single-cell genome-wide amplification method had a high failure rate of the established database of the embryo CNV-sequencing/SNP-sequencing techniques CNV/SNP sequencing.Conclusion:The MALBAC technique with more advantage is selected to build the technical platform of PGT of thalassemia successfully in southern Jiangxi area.

关 键 词：Α-地贫 Β-地贫 植入前胚胎遗传学诊断 单细胞全基因组扩增 MALBAC MDA 

分 类 号：R556.61[医药卫生—血液循环系统疾病]