UNASSIGNED: In this study, 68 blastocysts from preimplantation genetic testing (PGT)-assisted pregnancy were collected at our institution. On the fourth day of embryo culture, the embryos were transferred to a new medium for blastocyst culture and were washed either three times (NICS1 group) or ten times (NICS2 group). A trophectoderm (TE) biopsy was performed on the blastocysts, and the corresponding embryo culture media were collected for whole genome amplification (WGA) and high-throughput sequencing.
UNASSIGNED: The success rate of WGA was 100% (TE biopsy), 76.7% (NICS1 group), and 89.5% (NICS2 group). The success rate of WGA in embryo medium on days 5 and 6 of culture was 75.0% (33/44) and 100% (24/24), respectively. Using TE as the gold standard, the karyotype concordance rate between the results of the NICS1 and NICS2 groups\' embryo culture medium samples and TE results was 43.5% (10/23) and 73.5% (25/34), respectively. The sensitivity and specificity of detecting chromosomal abnormalities were higher in the NICS2 group than in the NICS1 group when TE was used (83.3% vs 60.0%; 62.5% vs 30.8%, respectively). The false-positive rate and false-negative rate (i.e., misdiagnosis rate and missed diagnosis rate, respectively) were lower in the NICS2 group than in the NICS1 group (37.5% vs 69.2%; 16.7% vs 40.0%, respectively).
UNASSIGNED: The NICS yielded favorable results after ten washings of the embryos. These findings provide a novel method for lowering the amount of cell-free DNA contamination from non-embryonic sources in the medium used for embryo development, optimizing the sampling procedure and improving the accuracy of the NICS test.
■在这项研究中,在我们的机构收集了来自植入前遗传测试(PGT)辅助妊娠的68个胚泡。胚胎培养的第四天,将胚胎转移到新的囊胚培养培养基中,洗涤3次(NICS1组)或10次(NICS2组).对胚泡进行滋养外胚层(TE)活检,收集相应的胚胎培养基进行全基因组扩增(WGA)和高通量测序。
■WGA的成功率为100%(TE活检),76.7%(NICS1组),和89.5%(NICS2组)。在培养的第5天和第6天,胚胎培养基中WGA的成功率分别为75.0%(33/44)和100%(24/24),分别。使用TE作为黄金标准,NICS1和NICS2组胚胎培养基样本与TE结果的核型一致率为43.5%(10/23)和73.5%(25/34),分别。使用TE时,NICS2组检测染色体异常的敏感性和特异性高于NICS1组(83.3%vs60.0%;62.5%vs30.8%,分别)。假阳性率和假阴性率(即误诊率和漏诊率,NICS2组分别低于NICS1组(37.5%vs69.2%;16.7%vs40.0%,分别)。
■NICS在胚胎洗涤十次后产生了良好的结果。这些发现提供了一种新颖的方法,可以降低用于胚胎发育的培养基中非胚胎来源的无细胞DNA污染量。优化采样程序,提高NICS测试的准确性。