单细胞微阵列比较基因组杂交技术在胚胎植入前遗传学筛查中的应用

摘要

目的 建立利用单细胞微阵列比较基因组杂交(array CGH,aCGH)技术进行胚胎植入前遗传学筛查(PGS)技术平台,并应用aCGH技术进行PGS获得持续妊娠. 方法 采集废弃胚胎来源卵裂球10个,核型正常男性(46,XY)淋巴细胞3个,通过单细胞全基因组扩增后,利用aCGH技术进行染色体非整倍体分析；在此基础上,将aCGH技术应用于临床一例反复流产患者. 结果 3个淋巴细胞及9个废胚来源卵裂球的全基因组扩增成功,1个废胚来源卵裂球扩增失败,扩增成功的样本通过aCGH检测后均获得明确的诊断,3个淋巴细胞分析结果与已知核型一致；临床病例5个胚胎活检的单卵裂球均得到明确诊断,移植1枚正常整倍体胚胎后获得临床持续妊娠. 结论 在胚胎植入前遗传学筛查中,单细胞aCGH技术能全面地评估胚胎染色体组情况,可应用于临床PGS.%Objective: To establish a reliable and accurate pre-implantation genetic screening (PGS) platform using single cell array comparative genomic hybridization (aCGH) technology.rnMethods: Ten blastomeres from discarded arrested embryos and 3 lymphocytes from a man with normal karyotype (46 XY) were amplified by whole genome amplification (WGA) and assessed by aCGH for 24 chromosomes. The validated aCGH technology was then applied for a couple who had previously experienced recurrent spontaneous abortions.rnResults: In preliminary tests, the whole genome amplification (WGA) product was successfully produced in 3 of 3 (100%) lymphocytes and 9 of 10 (90%) blastomeres. Following array CGH, the 3 lymphocytes showed the expected karyotype and all 9 blastomeres showed clear unambiguous aneuploidy chromosome profiles. In the first clinical case performed PGSj 2 of 5 embryos were found to be euploid. One euploid embryo was transferred back to the woman's uterus, which resulted in successful implantation and an ongoing pregnancy.rnConclusions: Single cell array CGH is a robust technology suitable for PGS. This is the first clinical successful case performed PGS using array CGH in mainland China.