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J Assist Reprod Genet. 2016 May;33(5):637-645. doi: 10.1007/s10815-016-0667-7. Epub 2016 Feb 22.

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

Author information

1
School of Medicine, Nankai University, Tianjin, 300071, China.
2
Department of Obstetrics and Gynecology, Chinese PLA General Hospital, Beijing, 100853, China.
3
Department of Obstetrics and Gynecology, Affiliated Hospital of Academy of Military Medical Sciences, Beijing, 100071, China.
4
Center for Reproductive Medicine, The First Hospital of Kunming, Kunming, 650011, China.
5
Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing, 100101, China.
6
Berry Genomics, Co., Ltd., Beijing, 100015, China.
7
Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing, 100101, China. sunbf@big.ac.cn.
8
Department of Obstetrics and Gynecology, Chinese PLA General Hospital, Beijing, 100853, China. yuanqingy@sina.cn.

Abstract

BACKGROUND:

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.

METHODS:

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).

RESULTS:

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 %.

CONCLUSION:

This study provides further evidence that BF is a promising alternative source of DNA for PGT.

KEYWORDS:

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

PMID:
26899834
PMCID:
PMC4870435
DOI:
10.1007/s10815-016-0667-7
[Indexed for MEDLINE]
Free PMC Article

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