使用经独立验证的下一代测序（NGS）技术对整倍体、非整倍体和嵌合体胚胎的滋养层活检进行重复性评估：一项初步研究。
The reproducibility of trophectoderm biopsies in euploid, aneuploid, and mosaic embryos using independently verified next-generation sequencing (NGS): a pilot study.
机构信息
NYU Langone Medical Center, 550 First Avenue, New York, NY, 10016, USA.
OC Fertility, 1401 Avocado Ave Ste 403, Newport Beach, CA, 92660-8725, USA.
出版信息
J Assist Reprod Genet. 2020 Mar;37(3):559-571. doi: 10.1007/s10815-020-01720-x. Epub 2020 Feb 28.
PURPOSE
To assess the accuracy and reliability of comprehensive chromosome screening by next-generation sequencing (NGS) of human trophectoderm (TE) biopsy specimens.
METHODS
The reliability and accuracy of diagnoses made by preimplantation genetic testing for aneuploidy (PGT-A) from TE biopsy were tested. Repeat biopsies of TE and inner cell mass (ICM) samples were obtained from thawed blastocysts previously tested by NGS. To test for the reliability of the NGS assay, biopsy samples were compared with the original PGT-A results. Prior NGS testing classified the TE samples as euploid, aneuploid, or aneuploid-mosaic. The resulting re-biopsied samples underwent SurePlex whole genome amplification followed by NGS via the MiSeq platform, with copy number value (CNV) determined using BlueFuse Multi Software. The primary outcome measure was reliability, defined as concordance between initial TE result and the repeat biopsies. Accuracy was determined by concordance between the TE and ICM samples, and compared between three chromosome types (disomic, aneuploid, and mosaic).
RESULTS
Re-biopsies were performed on 32 embryos with prior PGT-A showing euploidy (10 embryos), aneuploidy of one or two chromosomes (4 embryos), or aneuploid-mosaic with one aneuploid chromosome and one mosaic chromosome (18 embryos). One hundred twenty-nine biopsy samples completed NGS (90 TE and 39 ICM biopsies) and 105 biopsy results were included in the analysis. TE biopsies provide a highly accurate test of the future fetus, with the ICM disomic concordance rate of 97.6%. Clinical concordance rates indicate that TE biopsies provide a reliable test when the result is euploid (99.5%) or aneuploid (97.3%), but less reliable when the result is mosaic (35.2%).
CONCLUSION
TE biopsies predict euploidy or aneuploidy in the ICM with a high degree of accuracy. PGT-A with NGS of TE biopsies is shown to be highly reliable, with clinically relevant concordance rates for aneuploidy and euploidy over 95%. TE biopsies indicating mosaicism were less reliable (35.2%), presumably because mitotic non-disjunction events are not uniformly distributed throughout the blastocyst. However, classification of TE biopsy of PGT-A with NGS results as either aneuploid or euploid provides a highly reliable test.
目的
评估通过人类滋养外胚层（TE）活检标本进行下一代测序（NGS）的综合染色体筛查的准确性和可靠性。
方法
对胚胎植入前遗传学检测非整倍体（PGT-A）的诊断准确性和可靠性进行了测试。对先前通过 NGS 测试的解冻胚泡的 TE 和内细胞团（ICM）样本进行重复活检。为了测试 NGS 检测的可靠性，将活检样本与原始 PGT-A 结果进行比较。先前的 NGS 测试将 TE 样本分类为整倍体、非整倍体或非整倍体镶嵌。对重新活检的样本进行 SurePlex 全基因组扩增，然后通过 MiSeq 平台进行 NGS，使用 BlueFuse Multi 软件确定拷贝数值（CNV）。主要结局指标为初始 TE 结果与重复活检之间的一致性，定义为可靠性。通过 TE 和 ICM 样本之间的一致性来确定准确性，并在三种染色体类型（二倍体、非整倍体和镶嵌）之间进行比较。
结果
对先前 PGT-A 显示整倍体（10 个胚胎）、一两个染色体非整倍体（4 个胚胎）或一个非整倍体染色体和一个镶嵌染色体的非整倍体镶嵌（18 个胚胎）的 32 个胚胎进行了重复活检。完成了 129 个活检样本的 NGS（90 个 TE 和 39 个 ICM 活检样本），其中 105 个活检结果纳入分析。TE 活检对未来胎儿进行了高度准确的测试，ICM 二倍体的一致性率为 97.6%。临床一致性率表明，当结果为整倍体（99.5%）或非整倍体（97.3%）时，TE 活检是一种可靠的测试方法，但当结果为镶嵌体时，可靠性较低（35.2%）。
结论
TE 活检对 ICM 中的整倍体或非整倍体具有高度准确性。通过 TE 活检的 NGS 进行 PGT-A 被证明具有高度可靠性，非整倍体和整倍体的临床相关一致性率超过 95%。TE 活检显示镶嵌体的结果不太可靠（35.2%），可能是因为有丝分裂非分离事件在整个胚泡中不是均匀分布的。然而，将 NGS 结果的 PGT-A TE 活检分类为非整倍体或整倍体提供了一种高度可靠的测试方法。
Zotero 插件