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

Capsule:
Our study validated multiple annealing and looping-based amplification cycle next-generation sequencing for 24-chromosome aneuploidy screening of blastocysts.

Authors:
Jin Huang, Ph.D., Liying Yan, Ph.D., Sijia Lu, Ph.D., Nan Zhao, B.A., X. Sunney Xie, Ph.D., Jie Qiao, M.D.

Volume 105, Issue 6, Pages 1532-1536

Abstract:

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.

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