Massively Parallel Sequencing for Chromosomal Abnormality Testing in Trophectoderm Cells of Human Blastocysts
Massively Parallel Sequencing for Chromosomal Abnormality Testing in Trophectoderm Cells of Human Blastocysts
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DOI：
10.1095/biolreprod.112.106211
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发表时间：
2013-03-01
影响因子：
3.6
通讯作者：
Wang, Wei
中科院分区：
文献类型：
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作者：
Yin, XuYang;Tan, Ke;Wang, Wei
植入前的遗传学诊断和筛查被广泛用于染色体异常的识别，以避免移植有遗传缺陷的胚胎。大规模并行测序是一种快速发展的基因组分析方法，在临床上的应用越来越广泛。本研究的目的是利用MPS鉴定胚泡活检后的非整倍体和染色体不平衡重排。对16个体外受精周期的38个囊胚标本进行了滋养层胚层分析。使用Illumina HiSeq2000平台进行低覆盖率的全基因组测序，使用专门为染色体分析而创建的新算法。通过比较Affymetrix单核苷酸多态性(SNP)阵列获得的结果来评估这种MPS方法的效率。MPS检测到TE细胞的全基因组扩增产物，平均扩增深度为0.07倍，覆盖率为5.5%。整倍体胚胎26枚(68.4%)，均一非整倍体胚胎6枚(15.8%)。其中4个胚胎(10.5%)仅有染色体不平衡重排，其余两个胚胎(5.3%)既有非整倍体又有不平衡重排。除了一个样本外，几乎所有这些结果都得到了SNP阵列的证实，其中检测到了不同大小的不平衡重排，这可能是由于阵列分析中的染色体GC偏差所致。我们的研究表明，MPS可以以一种灵活且经济有效的策略和较高的潜在准确率来准确地检测胚胎染色体异常。
Preimplantation genetic diagnosis and screening are widely accepted for chromosomal abnormality identification to avoid transferring embryos with genetic defects. Massively parallel sequencing (MPS) is a rapidly developing approach for genome analysis with increasing application in clinical practice. The purpose of this study was to use MPS for identification of aneuploidies and unbalanced chromosomal rearrangements after blastocyst biopsy. Trophectoderm (TE) samples of 38 blastocysts from 16 in vitro fertilization cycles were subjected to analysis. Low-coverage whole genome sequencing was performed using the Illumina HiSeq2000 platform with a novel algorithm purposely created for chromosomal analysis. The efficiency of this MPS approach was estimated by comparing results obtained by an Affymetrix single-nucleotide polymorphism (SNP) array. Whole genome amplification (WGA) products of TE cells were detected by MPS, with an average of 0.07x depth and 5.5% coverage of the human genome. Twenty-six embryos (68.4%) were detected as euploid, while six embryos (15.8%) contained uniform aneuploidies. Four of these (10.5%) were with solely unbalanced chromosomal rearrangements, whereas the remaining two embryos (5.3%) showed both aneuploidies and unbalanced rearrangements. Almost all these results were confirmed by the SNP array, with the exception of one sample, where different sizes of unbalanced rearrangements were detected, possibly due to chromosomal GC bias in array analysis. Our study demonstrated MPS could be applied to accurately detect embryonic chromosomal abnormality with a flexible and cost-effective strategy and higher potential accuracy.