Foundation for Embryonic Competence, 140 Allen Road, Suite 300, Basking Ridge, NJ 07920, USA.
Reproductive Medicine Associates of New Jersey, 140 Allen Road, Basking Ridge, NJ 07920, USA.
Mol Hum Reprod. 2018 Jan 1;24(1):37-45. doi: 10.1093/molehr/gax060.
Can a novel targeted next generation sequencing (tNGS) platform accurately detect whole chromosome aneuploidy in a trophectoderm biopsy and provide additional information to improve testing?
Karyotypes obtained by tNGS were concordant with other validated platforms and single nucleotide polymorphism genotyping information obtained can be used for improved detection and quality control.
qPCR-based whole chromosome aneuploidy screening is highly accurate in comparison to other common methods and has been shown to improve IVF success in two randomized clinical trials. With aneuploidy screening becoming standard of care in many IVF centres, there is a need to develop platforms with high throughput, low cost capabilities.
STUDY DESIGN SIZE, DURATION: Twelve well-characterized cell lines were obtained from a commercial cell line repository and 31 discarded human blastocysts were obtained from 17 IVF patients who underwent comprehensive chromosome screening (CCS).
PARTICIPANTS/MATERIAL, SETTING, METHODS: All samples were processed using a unique amplification strategy which directly incorporated sequencing library adapters and barcodes. Sequencing was performed on an Ion Torrent Proton. A custom bioinformatics pipeline was used to determine the karyotype for each sample. The consistency of tNGS diagnoses with either conventional karyotyping of cell lines or quantitative real-time PCR based CCS of blastocyst biopsies was evaluated.
Overall consistency per sample of tNGS based CCS in 5-cell samples from a variety of cell lines was 99.2%. In the blinded analysis of rebiopsies of aneuploid blastocysts, an overall targeted tNGS CCS consistency of 98.7% was observed per sample. These data demonstrate the ability of tNGS based CCS to provide an accurate and high throughput evaluation of aneuploidy in the human blastocyst.
Not applicable.
This study is limited to whole chromosome aneuploidy, as mosaicism and segmental aneuploidy have not been investigated.
These data show an accurate, high throughput method, and with the greater depth of each amplicon sequenced in comparison to commercial kits, there is greater application available for single nucleotide polymorphism based analysis for quality control.
STUDY FUNDING/COMPETING INTERESTS: This study was funded through intramural research funds provided by the Foundation for Embryonic Competence. There are no competing interests.
新型靶向下一代测序（tNGS）平台能否准确检测滋养层活检中的全染色体非整倍体，并提供额外信息以改善检测？
通过 tNGS 获得的核型与其他经过验证的平台一致，并且可以使用获得的单核苷酸多态性基因分型信息来提高检测和质量控制的准确性。
与其他常见方法相比，基于 qPCR 的全染色体非整倍体筛查具有高度准确性，并且已经在两项随机临床试验中证明可以提高试管婴儿的成功率。随着非整倍体筛查成为许多试管婴儿中心的标准护理，因此需要开发具有高通量、低成本能力的平台。
研究设计、规模、持续时间：从商业细胞系库中获得了 12 个特征良好的细胞系，并从 17 名接受全面染色体筛查（CCS）的 IVF 患者中获得了 31 个废弃的人类囊胚。
参与者/材料、设置、方法：所有样本均采用独特的扩增策略进行处理，该策略直接包含测序文库接头和条形码。在 Ion Torrent Proton 上进行测序。使用定制的生物信息学管道来确定每个样本的核型。评估 tNGS 诊断与细胞系常规核型或囊胚活检基于定量实时 PCR 的 CCS 的一致性。
来自各种细胞系的 5 细胞样本中，基于 tNGS 的 CCS 的总体一致性为 99.2%。在对非整倍体囊胚再活检的盲法分析中，每个样本的总体靶向 tNGS CCS 一致性为 98.7%。这些数据表明，基于 tNGS 的 CCS 能够提供人类囊胚中染色体非整倍体的准确、高通量评估。
不适用。
局限性/谨慎原因：本研究仅限于全染色体非整倍体，因为未研究嵌合体和片段性非整倍体。
这些数据显示了一种准确、高通量的方法，并且与商业试剂盒相比，每个扩增子测序的深度更大，因此可以更广泛地应用于基于单核苷酸多态性的分析，以进行质量控制。
研究资金/竞争利益：本研究由胚胎能力基金会提供的内部研究资金资助。没有竞争利益。
