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...
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MDPI AG
2021-04-01
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| author | Anil Biricik Ettore Cotroneo Maria Giulia Minasi Pier Francesco Greco Sara Bono Matteo Surdo Federica Lecciso Mariateresa Sessa Francesco Fiorentino Francesca Spinella Ermanno Greco |
| author_facet | Anil Biricik Ettore Cotroneo Maria Giulia Minasi Pier Francesco Greco Sara Bono Matteo Surdo Federica Lecciso Mariateresa Sessa Francesco Fiorentino Francesca Spinella Ermanno Greco |
| author_sort | Anil Biricik |
| collection | DOAJ |
| description | 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<sup>TM</sup> Veriseq (Illumina, San Diego, CA, USA) and Ion Torrent Personal Genome Machine PGM<sup>TM</sup> 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. |
| first_indexed | 2024-03-10T12:25:23Z |
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| institution | Directory Open Access Journal |
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| last_indexed | 2024-03-10T12:25:23Z |
| publishDate | 2021-04-01 |
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| series | Life |
| spelling | doaj.art-b8ea219bf6904fb6b09ef3c998f8f6e12023-11-21T15:09:04ZengMDPI AGLife2075-17292021-04-0111434010.3390/life11040340Cross-Validation of Next-Generation Sequencing Technologies for Diagnosis of Chromosomal Mosaicism and Segmental Aneuploidies in Preimplantation Embryos ModelAnil Biricik0Ettore Cotroneo1Maria Giulia Minasi2Pier Francesco Greco3Sara Bono4Matteo Surdo5Federica Lecciso6Mariateresa Sessa7Francesco Fiorentino8Francesca Spinella9Ermanno Greco10Eurofins Genoma Group, Molecular Genetics Laboratories, Via Castel Giubileo 11, 00138 Rome, ItalyEurofins Genoma Group, Molecular Genetics Laboratories, Via Castel Giubileo 11, 00138 Rome, ItalyVilla Mafalda, Reproductive Medicine, 00199 Rome, ItalyVilla Mafalda, Reproductive Medicine, 00199 Rome, ItalyEurofins Genoma Group, Molecular Genetics Laboratories, Via Castel Giubileo 11, 00138 Rome, ItalyEurofins Genoma Group, Molecular Genetics Laboratories, Via Castel Giubileo 11, 00138 Rome, ItalyEurofins Genoma Group, Molecular Genetics Laboratories, Via Castel Giubileo 11, 00138 Rome, ItalyEurofins Genoma Group, Molecular Genetics Laboratories, Via Castel Giubileo 11, 00138 Rome, ItalyEurofins Genoma Group, Molecular Genetics Laboratories, Via Castel Giubileo 11, 00138 Rome, ItalyEurofins Genoma Group, Molecular Genetics Laboratories, Via Castel Giubileo 11, 00138 Rome, ItalyVilla Mafalda, Reproductive Medicine, 00199 Rome, ItalyDetection 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<sup>TM</sup> Veriseq (Illumina, San Diego, CA, USA) and Ion Torrent Personal Genome Machine PGM<sup>TM</sup> 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.https://www.mdpi.com/2075-1729/11/4/340next generation sequencingpreimplantation genetic testingchromosomal mosaicismmosaic embryossegmental aneuploidies |
| spellingShingle | Anil Biricik Ettore Cotroneo Maria Giulia Minasi Pier Francesco Greco Sara Bono Matteo Surdo Federica Lecciso Mariateresa Sessa Francesco Fiorentino Francesca Spinella Ermanno Greco Cross-Validation of Next-Generation Sequencing Technologies for Diagnosis of Chromosomal Mosaicism and Segmental Aneuploidies in Preimplantation Embryos Model Life next generation sequencing preimplantation genetic testing chromosomal mosaicism mosaic embryos segmental aneuploidies |
| title | Cross-Validation of Next-Generation Sequencing Technologies for Diagnosis of Chromosomal Mosaicism and Segmental Aneuploidies in Preimplantation Embryos Model |
| title_full | Cross-Validation of Next-Generation Sequencing Technologies for Diagnosis of Chromosomal Mosaicism and Segmental Aneuploidies in Preimplantation Embryos Model |
| title_fullStr | Cross-Validation of Next-Generation Sequencing Technologies for Diagnosis of Chromosomal Mosaicism and Segmental Aneuploidies in Preimplantation Embryos Model |
| title_full_unstemmed | Cross-Validation of Next-Generation Sequencing Technologies for Diagnosis of Chromosomal Mosaicism and Segmental Aneuploidies in Preimplantation Embryos Model |
| title_short | Cross-Validation of Next-Generation Sequencing Technologies for Diagnosis of Chromosomal Mosaicism and Segmental Aneuploidies in Preimplantation Embryos Model |
| title_sort | cross validation of next generation sequencing technologies for diagnosis of chromosomal mosaicism and segmental aneuploidies in preimplantation embryos model |
| topic | next generation sequencing preimplantation genetic testing chromosomal mosaicism mosaic embryos segmental aneuploidies |
| url | https://www.mdpi.com/2075-1729/11/4/340 |
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