Noninvasive preimplantation genetic testing for aneuploidy in spent medium may be more reliable than trophectoderm biopsy

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 116; no. 28; pp. 14105 - 14112
• Main Authors: Huang, Lei, Bogale, Berhan, Tang, Yaqiong, Lu, Sijia, Xie, Xiaoliang Sunney, Racowsky, Catherine
• Format: Journal Article
• Language: English
• Published: United States National Academy of Sciences 09.07.2019
• Series: PNAS Plus
• Subjects: Adult; Aneuploidy; Biological Sciences; Biopsy; Blastocyst - metabolism; Blastocyst - pathology; Blastocyst Inner Cell Mass - pathology; Blastocysts; Cell culture; Cell-Free Nucleic Acids - genetics; Chromosomes; Culture media; Culture Media - analysis; Deoxyribonucleic acid; DNA; Embryos; Female; Fertilization in Vitro - standards; Fetuses; Genetic screening; Genetic Testing; High-Throughput Nucleotide Sequencing; Humans; In vitro fertilization; Karyotype; Mosaicism; Next-generation sequencing; Noninvasive Prenatal Testing - standards; Ploidy; PNAS Plus; Pregnancy; Preimplantation Diagnosis - standards; Trophectoderm; human blastocyst; preimplantation genetic testing; ICSI; noninvasive PGT-A; culture medium
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.1907472116

Preimplantation genetic testing for aneuploidy (PGT-A) with trophectoderm (TE) biopsy is widely applied in in vitro fertilization (IVF) to identify aneuploid embryos. However, potential safety concerns regarding biopsy and restrictions to only those embryos suitable for biopsy pose limitations. In addition, embryo mosaicism gives rise to false positives and false negatives in PGT-A because the inner cell mass (ICM) cells, which give rise to the fetus, are not tested. Here, we report a critical examination of the efficacy of noninvasive preimplantation genetic testing for aneuploidy (niPGT-A) in the spent culture media of human blastocysts by analyzing the cell-free DNA, which reflects ploidy of both the TE and ICM. Fifty-two frozen donated blastocysts with TE biopsy results were thawed; each of their spent culture medium was collected after 24-h culture and analyzed by next-generation sequencing (NGS). niPGT-A and TE-biopsy PGT-A results were compared with the sequencing results of the corresponding embryos, which were taken as true results for aneuploidy reporting. With removal of all corona-cumulus cells, the false-negative rate (FNR) for niPGT-A was found to be zero. By applying an appropriate threshold for mosaicism, both the positive predictive value (PPV) and specificity for niPGT-A were much higher than TE-biopsy PGT-A. Furthermore, the concordance rates for both embryo ploidy and chromosome copy numbers were higher for niPGT-A than TE-biopsy PGT-A. These results suggest that niPGT-A is less prone to errors associated with embryo mosaicism and is more reliable than TE-biopsy PGT-A.