Diagnostic efficacy of blastocoel fluid and spent media as sources of DNA for preimplantation genetic testing in standard clinical conditions

Blastocoel fluid and spent blastocyst medium do not provide reliable DNA templates for minimally invasive or noninvasive preimplantation genetic testing when collected in standard clinical conditions.

0
0

Volume 110, Issue 5, Pages 870–879.e5

Authors:

Antonio Capalbo, Ph.D., Valeria Romanelli, Ph.D., Cristina Patassini, M.Sc., Maurizio Poli, D.Phil., Laura Girardi, M.Sc., Adriano Giancani, M.Sc., Marta Stoppa, M.Sc., Danilo Cimadomo, Ph.D., Filippo Maria Ubaldi, M.D., Laura Rienzi, M.Sc.

Abstract:

Objective

To determine whether blastocoel fluid (BF) or spent blastocyst medium (SBM) is a suitable template for genotype and/or karyotype assessment of in vitro fertilization–generated embryos.

Design

Prospective blinded study.

Setting

Genetic laboratory.

Patient(s)

From 26 patients undergoing preimplantation genetic testing (PGT) treatments, 103 trophectoderms (TE), 92 BF samples, and 72 SBM samples.

Intervention(s)

The BF and SBM were retrieved at the time of TE biopsy. Two DNA extraction strategies were evaluated on independent BF and SBM samples. Further enrolled samples were processed using next-generation sequencing and quantitative polymerase chain reaction for assessment of monogenic disorders (PGT-M) or aneuploidy (PGT-A).

Main Outcome Measure(s)

DNA amplification and concordance rates across BF, SBM, and TE to assess diagnostic efficiency.

Result(s)

No differences were detected among the DNA extraction methods tested. In PGT-M tests, for BF and SBM, 2.9% and 20.8% of all samples, respectively, produced a diagnosis concordant with the corresponding TE (n = 2 of 69 and 15 of 72, respectively). The SBM samples were associated with higher discordance rates and higher artifacts/contamination detection compared with BF. In multiple occasions, the maternal mutated variant was detected in the SBM of homozygous wild-type embryos, showing evidence of maternal DNA persistence in culture medium. In PGT-A tests, BF analysis showed high amplification failure rates (65.2%) and an overall concordance rate of 37.5% among amplified samples.

Conclusion(s)

Based on current methodologies, BF and SBM genetic analyses do not provide sufficiently reliable results to be employed clinically. Until the risk of maternal contamination can be properly prevented, SBM should not be used for PGT-M purposes.


Read the full text here.

Medium untitled 1

Fertility and Sterility

Editorial Office, American Society for Reproductive Medicine

Fertility and Sterility® is an international journal for obstetricians, gynecologists, reproductive endocrinologists, urologists, basic scientists and others who treat and investigate problems of infertility and human reproductive disorders. The journal publishes juried original scientific articles in clinical and laboratory research relevant to reproductive endocrinology, urology, andrology, physiology, immunology, genetics, contraception, and menopause. Fertility and Sterility® encourages and supports meaningful basic and clinical research, and facilitates and promotes excellence in professional education, in the field of reproductive medicine.

No comments yet.