Molecular analysis of DNA in blastocoele fluid using next-generation sequencing

作者：Zhang, Yixin; Li, Na; Wang, Li; Sun, Huiying; Ma, Minyue; Wang, Hui; Xu, Xiaofei; Zhang, Wenke; Liu, Yingyu; Cram, David S.; Sun, Baofa^*; Yao, Yuanqing^*

来源：Journal of Assisted Reproduction and Genetics, 2016, 33(5): 637-645.

DOI：10.1007/s10815-016-0667-7

• Blastocoele fluid
• Blastomere
• Next-generation sequencing
• Preimplantation genetic testing
• Bioinformatics

Preimplantation genetic testing (PGT) requires an invasive biopsy to obtain embryonic material for genetic analysis. The availability of a less invasive procedure would increase the overall efficacy of PGT. The aim of the study was to explore the potential of blastocoele fluid (BF) as an alternative source of embryonic DNA for PGT.
Collection of BF was performed by aspiration with a fine needle prior to vitrification. BF DNA was subjected to whole-genome amplification (WGA) and analyzed by high-resolution next-generation sequencing (NGS).
A high-quality WGA product was obtained from 8 of 11 (72.7 %) samples. Comparison of matching BF and blastomere samples showed that the genomic representation of sequencing reads was consistently similar with respect to density and regional coverage across the 24 chromosomes. A genome-wide survey of the sample sequencing data also indicated that BF was highly representative of known single gene sequences, and this observation was validated by PCR analyses of ten randomly selected genes, with an overall efficiency of 84 %.
This study provides further evidence that BF is a promising alternative source of DNA for PGT.

• 出版日期2016-5
• 单位中国人民解放军总医院; 昆明市第一人民医院; 中国人民解放军军事医学科学院; 南开大学; 中国科学院北京基因组研究所