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

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., and Laura Rienzi, M.Sc.

Fertility and Sterility® Vol. 110, No. 5, October 2018 0015-0282/

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): TheBFandSBMwereretrievedatthetimeofTEbiopsy.TwoDNAextractionstrategieswereevaluatedonindependent 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): NodifferencesweredetectedamongtheDNAextractionmethodstested. In PGT Mtests, for BF and SBM, 2.9% and 20.8% of all samples, respectively, produced a diagnosis concordant with the corresponding TE (n 1⁄4 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. (Fertil Steril! 2018;110:870–9. !2018 by American Society for Reproductive Medicine.)
El resumen está disponible en Español al final del artículo.

Key Words: Blastocentesis, blastocoel, noninvasive, PGT, spent blastocyst media