Genomic DNA in human blastocoele fluid
Genomic DNA in human blastocoele fluid
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DOI：
10.1016/j.rbmo.2013.02.012
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发表时间：
2013-06-01
影响因子：
4
通讯作者：
Bulletti, C.
中科院分区：
文献类型：
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作者：
Palini, S.;Galluzzi, L.;Bulletti, C.
体外受精通常需要通过玻璃化冷冻保存胚胎。在玻璃化过程中，可以通过抽出囊胚腔液体来使胚胎塌陷。最近通过高效液相色谱-电喷雾电离-质谱法研究了囊胚腔液的代谢组学特征，以提供有助于估计植入效率的代谢物信息。然而，迄今为止尚未报道囊胚腔液中存在胚胎DNA。这项研究表明，使用实时 PCR 技术，在玻璃化冷冻过程中收获的囊胚腔液体样本中，约 90% 都存在基因组 DNA。此外，通过扩增多拷贝基因 TSPY1（位于 Y 染色体上）和 TBC1D3（位于 17 号染色体上），证明了直接从囊胚腔液中确定胚胎性别的潜力。这开启了从携带 X 连锁疾病的夫妇中筛选胚胎以识别具有高患病风险的男性胚胎的可能性。全基因组扩增技术在液体样本中的应用也被证明是可行的，有可能允许更全面的基因测试。作为原理证明，尝试使用微阵列比较基因组杂交来确认胚胎的性别并检测几种非整倍体。然而，需要进一步的研究来验证这种方法并确认其准确性足以用于诊断目的。 (C) 2013 年，Reproductive Healthcare Ltd. 由 Elsevier Ltd 出版。保留所有权利。
IVF often requires embryo cryopreservation through vitrification. During the vitrification process, the embryos can be collapsed by withdrawing the blastocoele fluid. The metabolomic profile of blastocoele fluid has been recently investigated by high-performance liquid chromatography-electrospray ionization-mass spectrometry to provide metabolite information that can help estimations of implantation efficiency. However, the presence of embryo DNA in blastocoele fluid has not been reported to date. This study shows using real-time PCR that genomic DNA was present in about 90% of blastocoele fluid samples harvested during the vitrification procedure. Moreover, the potential for determining embryo sex directly from blastocoele fluid is demonstrated by amplifying the multicopy genes TSPY1 (on the Y chromosome) and TBC1D3 (on chromosome 17). This opens up the possibility of screening embryos from couples carrying an X-linked disorder to identify male embryos at high risk of disease. The application of whole-genome amplification technologies to fluid samples is also shown to be feasible, potentially allowing more comprehensive genetic tests. As proof of principle, microarray comparative genomic hybridization was attempted to confirm the sex of embryos as well as detect several aneuploidies. However, further studies are needed to validate this approach and confirm that the accuracy is sufficient for diagnostic purposes. (C) 2013, Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.