Validation of a next-generation sequencing–based protocol for 24-chromosome aneuploidy screening of blastocysts

Objective To validate a 24-chromosome aneuploidy preimplantation genetic screening protocol based on multiple annealing and looping-based amplification cycle (MALBAC) and next-generation sequencing (NGS). Design Single-nucleotide polymorphism (SNP) array and MALBAC-NGS analysis. Setting University-affiliated in vitro fertilization (IVF) center. Patient(s) Fifteen women from whom 30 blastocysts were obtained for genotyping. Intervention(s) Not applicable. Main Outcome Measure(s) Chromosomal status comparison of results of array comparative genomic hybridization (aCGH), SNP array, and MALBAC-NGS for 24-chromosome aneuploidy screening. Result(s) Trophectoderm biopsy samples from blastocysts were first analyzed using array comparative genomic hybridization (aCGH); the embryos with detected with chromosomal abnormalities were rebiopsied, and dissociated into two portions, and subjected to SNP array and MALBAC-NGS for 24-chromosome aneuploidy screening. All 30 samples were successfully genotyped by array CGH, SNP array, and MALBAC-NGS. All blastocysts were correctly identified as aneuploid, and there was a 100% concordance in terms of diagnosis provided between the three methods. In the 720 detected chromosomes, the concordance rate between MALBAC-NGS and array CGH was 99.31% (715 of 720), and the concordance rate between MALBAC-NGS and SNP array was 99.58% (717 of 720). When compared with aCGH, MALBAC-NGS specificity for aneuploidy call was 99.85% (674 of 675; 95% CI, 99.17–99.97) with a sensitivity of 91.11% (41 of 45; 95% CI, 79.27–96.49). When compared with SNP array, MALBAC-NGS specificity for aneuploidy call was 99.85% (676 of 677; 95% CI, 99.17–99.97) with a sensitivity of 95.35% (41 of 43; 95% CI, 85.54–98.72). Conclusion(s) MALBAC-NGS provides concordant chromosomal results when compared with aCGH and SNP array in blastocysts with chromosomal abnormalities.