【摘要】 目的 探讨利用囊胚培养液中游离DNA检测胚胎整倍性的样品留取优化方案。方法 本研究共纳入239枚囊胚，所有的囊胚均来源于北京大学第三医院生殖中心胚胎植入前遗传学整倍体检测技术(PGT-A)周期。纳入患者的PGT-A指征为女方高龄(>38岁)、复发性流产、反复着床失败。胚胎在D3天更换囊胚培养液，并采取单胚胎单独培养，收集可以PGT-A活检囊胚的培养液。收集方法根据是否胚胎打孔以及胚胎培养时长分为三个阶段。PGT-A囊胚活检的滋养层细胞采用SNP芯片检测整倍性；培养液游离DNA采用二代测序方法(NGS)进行无创胚胎染色体整倍性筛查(NiPGT-A);背靠背比较两者的结果，分析NiPGT-A方法的检出率、NiPGT-A结果与PGT-A结果倍性一致性、整倍体符合率、非整倍体符合率、颗粒细胞污染率等。共有56枚PGT-A整倍性胚胎移植，分析对应胚胎的NiPGT-A结果，统计临床妊娠结局。结果 239枚囊胚中209枚囊胚有NiPGT-A结果，检出率为87.5%。随着采样方法的改进，NiPGT-A结果与PGT-A结果倍性一致性、整倍体符合率逐渐提高；颗粒细胞污染率逐渐降低。239枚胚胎中有56枚PGT-A整倍体的胚胎移植，23枚胚胎的结局为活产/持续妊娠。NiPGT-A没有结果的胚胎妊娠率较高(71.4%),高于NiPGT-A为整倍体或者非整倍体胚胎的临床妊娠结局(33.3%,40.9%)。结论 囊胚培养液的样品留取方法对于NiPGT-A检测至关重要，推荐D3天再次去除颗粒细胞、采集D4-D5/6天囊胚培养液，而且不要针对胚胎进行打孔处理。更多还原

【Abstract】 Objective To explore the feasibility of noninvasive preimplantation genetic testing for aneuploidy(NiPGT-A) and the optimization method of sample retention. Methods 239 PGT-A blastocysts from Center for Reproductive Medicine of Peking University Third Hospital were included. The PGA-A indications of included women were older age(>38 years), recurrent abortion, and repeated implantation failure. If the blastocyst was up to the biopsy standard, the corresponding culture medium was collected. The collection method was divided into three stages according to embryo drilling and the culture duration. The trophectoderm cells were detected by SNP array, while NiPGT-A was detected by next generation sequencing(NGS). PGT-A and NiPGT-A results including the detection rate of NiPGT-A, the consistency of NiPGT-A and PGT-A results, the coincidence rate of aneuploidy and granulosa cell pollution rate, were compared back-to-back. Only 56 PGT-A euploid embryos were transferred and the NiPGT-A results of the transferred blastocysts were analyzed. Results Among 239 blastocysts, 209 blastocysts had NiPGT-A results, and the detection rate of NiPGT-A was 87.5%. With the improvement of sampling method, the consistency of ploidy and aneuploidy of NiPGT-A results and PGT-A results ewere gradually increased; the pollution rate of granular cells was gradually reduced. Among 56 PGT-A euploidy blastocysts transferred, 23 blastocysts got live births or ongoing pregnancies. However, the clinical pregnancy rate was 71.4% in the group of the embryos without NiPGT-A result, higher than that of the group of NiPGT-A euploidy(33.3%) and the group of NiPGT-A aneuploidy(40.9%). Conclusion It is recommended to collect blastocyst culture medium from D4 to D5/D6, without drilling the embryos. With continuous improvement of NiPGT-A technology, it is expected to be used in clinical practice.更多还原