María Vera-Rodríguez1, Claude-Edouard Michel2, Amparo Mercader3, Alex J Bladon2, Lorena Rodrigo4, Felix Kokocinski2, Emilia Mateu4, Nasser Al-Asmar5, David Blesa4, Carlos Simón6, Carmen Rubio4. 1. Igenomix, Valencia, Spain. Electronic address: maria.vera@igenomix.com. 2. Illumina, Cambridge, United Kingdom. 3. Instituto Universitario IVI, Valencia University, Spain; Fundación Instituto Valenciano de Infertilidad (FIVI)/INCLIVA, Valencia, Spain. 4. Igenomix, Valencia, Spain; Fundación Instituto Valenciano de Infertilidad (FIVI)/INCLIVA, Valencia, Spain. 5. IviGen, Miami, Florida. 6. Igenomix, Valencia, Spain; Instituto Universitario IVI, Valencia University, Spain; Fundación Instituto Valenciano de Infertilidad (FIVI)/INCLIVA, Valencia, Spain; Department of Obstetrics and Gynecology, School of Medicine, Stanford University, Stanford, California.
Abstract
OBJECTIVE: To evaluate the ability of next-generation sequencing (NGS) to detect pure and mosaic segmental aneuploidies in trophectoderm biopsies and to identify distribution patterns in whole blastocysts. DESIGN: Validation study. SETTING: Reference laboratory. PATIENT(S): Seventy couples with known karyotypes who had undergone preimplantation genetic screening with diagnoses at the blastocyst stage using array comparative genomic hybridization (aCGH). INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Concordance rates for segmental and whole-chromosome aneuploidies determined between aCGH and NGS, and estimates of mosaicism levels of segmental aneuploidies in fixed blastocysts. RESULT(S): We used NGS with amplified DNA from trophectoderm biopsies in which segmental aneuploidies had been previously detected by array comparative genomic hybridization (aCGH). Single-cell fluorescent in situ hybridization (FISH) was then used as an independent form of analysis. The concordance rate between NGS and aCGH was 124 (98.4%) of 126 for the detection of segmental aneuploidies, and 48 (96.0%) of 50 for whole-chromosome aneuploidies. The overall concordance rate was 99.8% (2,276 of 2,280 chromosomes assessed). After FISH analyses with 41.4 ± 24.3 cells per blastocyst, 26 (92.9%) of 28 segmentals detected by aCGH and NGS were confirmed. The FISH analysis did not detect the segmentals in two blastocysts, in which all cells analyzed were euploid. CONCLUSION(S): This is the first report analyzing distribution patterns of segmental aneuploidies in trophectoderm biopsy by NGS. We have demonstrated that NGS allows the detection of pure and mosaic segmental aneuploidies with the same efficiency as aCGH. The FISH analysis confirmed the existence of these events in the trophectoderm and the inner cell mass.
OBJECTIVE: To evaluate the ability of next-generation sequencing (NGS) to detect pure and mosaic segmental aneuploidies in trophectoderm biopsies and to identify distribution patterns in whole blastocysts. DESIGN: Validation study. SETTING: Reference laboratory. PATIENT(S): Seventy couples with known karyotypes who had undergone preimplantation genetic screening with diagnoses at the blastocyst stage using array comparative genomic hybridization (aCGH). INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Concordance rates for segmental and whole-chromosome aneuploidies determined between aCGH and NGS, and estimates of mosaicism levels of segmental aneuploidies in fixed blastocysts. RESULT(S): We used NGS with amplified DNA from trophectoderm biopsies in which segmental aneuploidies had been previously detected by array comparative genomic hybridization (aCGH). Single-cell fluorescent in situ hybridization (FISH) was then used as an independent form of analysis. The concordance rate between NGS and aCGH was 124 (98.4%) of 126 for the detection of segmental aneuploidies, and 48 (96.0%) of 50 for whole-chromosome aneuploidies. The overall concordance rate was 99.8% (2,276 of 2,280 chromosomes assessed). After FISH analyses with 41.4 ± 24.3 cells per blastocyst, 26 (92.9%) of 28 segmentals detected by aCGH and NGS were confirmed. The FISH analysis did not detect the segmentals in two blastocysts, in which all cells analyzed were euploid. CONCLUSION(S): This is the first report analyzing distribution patterns of segmental aneuploidies in trophectoderm biopsy by NGS. We have demonstrated that NGS allows the detection of pure and mosaic segmental aneuploidies with the same efficiency as aCGH. The FISH analysis confirmed the existence of these events in the trophectoderm and the inner cell mass.
Authors: Laura Girardi; Munevver Serdarogullari; Cristina Patassini; Maurizio Poli; Marco Fabiani; Silvia Caroselli; Onder Coban; Necati Findikli; Fazilet Kubra Boynukalin; Mustafa Bahceci; Rupali Chopra; Rita Canipari; Danilo Cimadomo; Laura Rienzi; Filippo Ubaldi; Eva Hoffmann; Carmen Rubio; Carlos Simon; Antonio Capalbo Journal: Am J Hum Genet Date: 2020-03-26 Impact factor: 11.025
Authors: Karen Sermon; Antonio Capalbo; Jacques Cohen; Edith Coonen; Martine De Rycke; Anick De Vos; Joy Delhanty; Francesco Fiorentino; Norbert Gleicher; Georg Griesinger; Jamie Grifo; Alan Handyside; Joyce Harper; Georgia Kokkali; Sebastiaan Mastenbroek; David Meldrum; Marcos Meseguer; Markus Montag; Santiago Munné; Laura Rienzi; Carmen Rubio; Katherine Scott; Richard Scott; Carlos Simon; Jason Swain; Nathan Treff; Filippo Ubaldi; Rita Vassena; Joris Robert Vermeesch; Willem Verpoest; Dagan Wells; Joep Geraedts Journal: Mol Hum Reprod Date: 2016-06-02 Impact factor: 4.025