目的:为了确定使用基于下一代测序(NGS)的非整倍体植入前遗传学检测(PGT-A)测试的单个滋养外胚层活检结果的非整倍体结果与使用基于单核苷酸多态性(SNP)阵列的PGT-A平台测试的再活检结果是否一致。
方法:盲目前瞻性队列研究。
方法:大学附属生育中心。
方法:从捐赠的样本中选择100个胚泡;在基于NGS的PGT-A的滋养外胚层活检中,40只具有至少一个全染色体全拷贝数非整倍性,20具有单个全染色体中间拷贝数(“全染色体镶嵌”),20个有一个完整的片段非整倍体,和20具有单个片段中间拷贝数(“片段马赛克”)。
方法:从每个胚胎收集四次再活检:三次滋养外胚层活检和剩余的胚胎。每次再活检都是随机的,失明,并使用基于SNP阵列的PGT-A平台进行评估,该平台结合了拷贝数和等位基因比率分析,没有镶嵌主义报道。
方法:一致性:1)NGS结果和再活检结果之间,和2)在每个胚胎的盲检结果中。
结果:在95%(95%置信区间[CI]83-99%)的胚胎中重新确认了NGS诊断的全染色体非整倍体;在所有确凿的活检中,两个具有NGS诊断的全染色体非整倍体的胚胎被称为整倍体。在诊断为NGS的全染色体镶嵌的胚胎中,在所有结论性的再活检中,35%(95%CI15-59%)被称为整倍体,15%(95%CI3-38%)被称为全染色体非整倍体。在所有结论性的重新活检中,有30%(95%CI12-54%)的NGS诊断的节段性非整倍体的胚胎和65%(95%CI41-85%)的NGS诊断的节段性镶嵌性的胚胎被称为整倍体。总的来说,13%(95%CI6-25%)的具有NGS诊断的全拷贝数非整倍体的胚胎和50%(95%CI34-66%)的具有NGS诊断的镶嵌性的胚胎具有一致的整倍体SNP结果。相反,具有至少一个非整倍体SNP结果(n=72)的所有胚胎均具有:1)在同一胚胎的另一次活检中,SNP诊断为非整倍体,或2)涉及相同染色体的NGS诊断的非整倍性/镶嵌性。
结论:NGS诊断的全染色体非整倍体与使用基于SNP阵列的PGT-A测试的再活检高度一致;然而,全染色体镶嵌,节段性非整倍性,分段镶嵌不那么一致,增强具有这些结果的胚胎可能具有生殖潜力并适合转移。
OBJECTIVE: To determine how often a noneuploid result from a single trophectoderm (TE) biopsy tested with the next-generation sequencing (NGS)-based preimplantation genetic testing for aneuploidy (PGT-A) is concordant with rebiopsies tested with a single-nucleotide polymorphism (SNP) array-based PGT-A platform.
METHODS: Blinded prospective cohort study.
METHODS: University-affiliated fertility center.
METHODS: One hundred blastocysts were chosen from donated samples; on TE biopsy with NGS-based PGT-A, 40 had at least one whole chromosome full copy number aneuploidy alone, 20 had a single whole chromosome intermediate copy number (\"whole chromosome mosaic\"), 20 had a single full
segmental aneuploidy (segA), and 20 had a single
segmental intermediate copy number (\"
segmental mosaic\").
METHODS: Four rebiopsies were collected from each embryo: 3 TE biopsies and the remaining embryo. Each rebiopsy was randomized, blinded, and assessed with an SNP array-based PGT-A platform that combines copy number and allele ratio analyses, without mosaicism reporting.
METHODS: Concordance between the NGS result and rebiopsy results and within each embryo\'s blinded rebiopsy results.
RESULTS: Next-generation sequencing-diagnosed whole chromosome aneuploidy (WCA) was reconfirmed in 95% (95% confidence interval [CI], 83%-99%) of embryos; 2 embryos with NGS-diagnosed WCA were called euploid on all conclusive rebiopsies. Among embryos with NGS-diagnosed whole chromosome mosaicism, 35% (95% CI, 15%-59%) were called euploid and 15% (95% CI, 3%-38%) were called whole chromosome aneuploid on all conclusive rebiopsies. A total of 30% (95% CI, 12%-54%) of embryos with NGS-diagnosed segA and 65% (95% CI, 41%-85%) of embryos with NGS-diagnosed
segmental mosaicism were called euploid on all conclusive rebiopsies. In total, 13% (95% CI, 6%-25%) of embryos with NGS-diagnosed full copy number aneuploidy and 50% (95% CI, 34%-66%) of embryos with NGS-diagnosed mosaicism had uniformly euploid SNP results. Conversely, all embryos with at least one noneuploid SNP result (n = 72) either had SNP-diagnosed aneuploidy on another rebiopsy from the same embryo or NGS-diagnosed aneuploidy/mosaicism involving the same chromosome.
CONCLUSIONS: Next-generation sequencing-diagnosed WCA is highly concordant with rebiopsies tested with an SNP array-based PGT-A; however, whole chromosome mosaicism, segA, and
segmental mosaicism are less concordant, reinforcing that embryos with these results may have reproductive potential and be suitable for transfer.