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Semiconductor Sequencing for Preimplantation Genetic Testing for Aneuploidy
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Preimplantation High-Resolution HLA Sequencing Using Next Generation Sequencing.

Maryam Rafati1, Mohammad Mahdi Akhondi2, Mohammad Reza Sadeghi2

  • 1Reproductive Biotechnology Research Center, Avicenna Research Institute, Academic Center for Education, Culture and Research (ACECR), Tehran, Iran; Fetal Health Research Center, Hope Generation Foundation, Tehran, Iran; Gene Clinic, Tehran, Iran.

Biology of Blood and Marrow Transplantation : Journal of the American Society for Blood and Marrow Transplantation
|April 13, 2018
PubMed
Summary

Finding a matched donor for hematopoietic stem cell transplantation (SCT) is challenging. This study shows preimplantation human leukocyte antigen (HLA) sequencing using next-generation sequencing (NGS) is feasible for identifying HLA-identical embryos.

Keywords:
Hematopoietic stem cell transplantationHigh-resolution HLA typingMultiple displacement amplificationNext generation sequencingPreimplantation genetic diagnosis

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Area of Science:

  • Genetics
  • Reproductive Medicine
  • Immunology

Background:

  • Hematopoietic stem cell transplantation (SCT) is crucial for treating hematologic disorders and cancers.
  • Finding an HLA-identical donor is often difficult, leaving families with limited options.
  • Conceiving a "savior baby" is sometimes pursued, especially where donor registries are unavailable.

Purpose of the Study:

  • To assess the feasibility of preimplantation human leukocyte antigen (HLA) sequencing on single blastomeres using next-generation sequencing (NGS).
  • To evaluate a novel approach for identifying HLA-matched embryos for families needing SCT.

Main Methods:

  • Embryos from two couples undergoing preimplantation genetic diagnosis for β-thalassemia were analyzed.
  • High-resolution HLA typing of 5 loci was performed on single cells using multiple displacement amplification and NGS.
  • Analysis included assessing amplification efficiency, allele dropout (ADO) rates, and resolving ambiguities through haplotype analysis.

Main Results:

  • Conclusive 4-digit HLA typing was achieved for 88.9% of 90 HLA alleles.
  • High amplification efficiency (93.3%) was observed, with an allele dropout rate of 22.2%.
  • Minor ambiguities in allelic and protein levels were resolved using haplotype analysis.

Conclusions:

  • Preimplantation HLA sequencing using NGS is a feasible technique.
  • This method combines single-cell whole genome amplification, preimplantation genetic diagnosis, and high-resolution HLA typing.
  • This approach offers a practical solution for families seeking an HLA-matched donor for SCT.