Validation of next-generation sequencing for comprehensive chromosome screening of embryos
Validation of next-generation sequencing for comprehensive chromosome screening of embryos
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DOI：
10.1016/j.rbmo.2015.09.002
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发表时间：
2015-12-01
影响因子：
4
通讯作者：
Wells, Dagan
中科院分区：
文献类型：
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作者：
Kung, Allen;Munne, Santiago;Wells, Dagan
大规模并行基因组测序，也称为下一代测序（NGS），是植入前遗传学诊断的最新方法。本研究的目的是确定NGS是否可以准确地检测人类胚胎的非整倍体。低覆盖率基因组测序应用于发育的胚泡阶段的胚胎的滋养外胚层活检。通过将结果与先前验证的平台阵列比较基因组杂交（aCGH）进行比较，确定NGS的灵敏度和特异性。共计156个样本（116个进行盲法评估）进行了测试：40个样品是胚泡的再活检，其中原始活检标本先前进行了aCGH测试; 4个样品是来自先前在卵裂期活检并使用aCGH测试的胚胎的单卵裂球的再活检; 18个样品是来源于良好表征的细胞系的单细胞; 94个样本是来自胚胎活检的全基因组扩增产物，这些胚胎活检是使用aCGH分析的先前植入前遗传筛查周期。每个胚胎，NGS灵敏度为100%（无假阴性），特异性为100%（无假阳性）。每条染色体的NGS一致率为99.20%。随着更多的改进，NGS将允许同时诊断单基因疾病和非整倍性，并可能有潜力提供更详细的了解胚胎活力的其他方面。(C)2015生殖保健有限公司由爱思唯尔有限公司出版。版权所有。
Massively parallel genome sequencing, also known as next-generation sequencing (NGS), is the latest approach for preimplantation genetic diagnosis. The purpose of this study was to determine whether NGS can accurately detect aneuploidy in human embryos. Low coverage genome sequencing was applied to trophectoderm biopsies of embryos at the blastocyst stage of development. Sensitivity and specificity of NGS was determined by comparison of results with a previously validated platform, array-comparative genomic hybridization (aCGH). In total, 156 samples (116 were blindly assessed) were tested: 40 samples were re-biopsies of blastocysts where the original biopsy specimen was previously tested for aCGH; four samples were re-biopsies of single blastomeres from embryos previously biopsied at the cleavage stage and tested using aCGH; 18 samples were single cells derived from well-characterized cell lines; 94 samples were whole-genome amplification products from embryo biopsies taken from previous preimplantation genetic screening cycles analysed using aCGH. Per embryo, NGS sensitivity was 100% (no false negatives), and 100% specificity (no false positives). Per chromosome, NGS concordance was 99.20%. With more improvement, NGS will allow the simultaneous diagnosis of single gene disorders and aneuploidy, and may have the potential to provide more detailed insight into other aspects of embryo viability. (C) 2015 Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.