Cross-Validation of Next-Generation Sequencing Technologies for Diagnosis of Chromosomal Mosaicism and Segmental Aneuploidies in Preimplantation Embryos Model

Detection of mosaic embryos is crucial to offer more possibilities of success to women undergoing in vitro fertilization (IVF) treatment. Next Generation Sequencing (NGS)-based preimplantation genetic testing are increasingly used for this purpose since their higher capability to detect chromosomal mosaicism in human embryos. In the recent years, new NGS systems were released, however their performance for chromosomal mosaicism are variable. We performed a cross-validation analysis of two different NGS platforms in order to assess the feasibility of these techniques and provide standard parameters for the detection of such aneuploidies. The study evaluated the performance of Miseq(TM) Veriseq (Illumina, San Diego, CA, USA) and Ion Torrent Personal Genome Machine PGM(TM) ReproSeq (Thermo Fisher, Waltham, MA, USA) for the detection of whole and segmental mosaic aneuploidies. Reconstructed samples with known percentage of mosaicism were analyzed with both platforms and sensitivity and specificity were determined. Both platforms had high level of specificity and sensitivity with a Limit Of Detection (LOD) at >= 30% of mosaicism and a showed a >= 5.0 Mb resolution for segmental abnormalities. Our findings demonstrated that NGS methodologies are capable of accurately detecting chromosomal mosaicism and segmental aneuploidies. The knowledge of LOD for each NGS platform has the potential to reduce false-negative and false-positive diagnoses when applied to detect chromosomal mosaicism in a clinical setting.

Automated summary

The study evaluated the performance of two different NGS platforms for the detection of whole and segmental mosaic aneuploidies, demonstrating high specificity and sensitivity in detecting mosaicism above 30% and a resolution of segmental abnormalities above 5.0 Mb. The findings suggest that NGS methodologies are capable of accurately detecting chromosomal mosaicism and segmental aneuploidies, potentially reducing false-negative and false-positive diagnoses in a clinical setting.