Novel Multiplex Fluorescent PCR-Based Method for HLA Typing and Preimplantational Genetic Diagnosis of β-Thalassemia

Background and Aims Thalassemia is curable by bone marrow transplantation; however, finding suitable donors with defined HLA combination remains a major challenge. Cord blood stem cells with preselected HLA system through preimplantation genetic diagnosis (PGD) proved very useful for resolving scarc...

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Bibliographic Details
Published in:Archives of medical research 2016-05, Vol.47 (4), p.293-298
Main Authors: Khosravi, Sharifeh, Salehi, Mansour, Ramezanzadeh, Mahboobeh, Mirzaei, Hamed, Salehi, Rasoul
Format: Article
Language:English
Subjects:
beta-Globins - genetics
beta-Thalassemia - genetics
beta-Thalassemia - immunology
Female
Fertilization in Vitro
Fluorescence
Genetic Loci
Histocompatibility Testing - methods
HLA typing
Humans
Internal Medicine
Multiplex fluorescent PCR
Multiplex Polymerase Chain Reaction
Mutation
Pregnancy
Preimplantation Diagnosis - methods
Preimplantation genetic diagnosis
STR markers
β-thalassemia
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Summary:Background and Aims Thalassemia is curable by bone marrow transplantation; however, finding suitable donors with defined HLA combination remains a major challenge. Cord blood stem cells with preselected HLA system through preimplantation genetic diagnosis (PGD) proved very useful for resolving scarce HLA-matched bone marrow donors. Methods A thalassemia trait couple with an affected child was included in this study. We used informative STR markers at the HLA and beta globin loci to develop a single cell multiplex fluorescent PCR protocol. The protocol was extensively optimized on single lymphocytes isolated from the couple's peripheral blood. The optimized protocol was applied on single blastomeres biopsied from day 3 cleavage stage IVF embryos of the couple. Results Four IVF embryos biopsied on day 3 and a single blastomere of each were provided for genetic diagnosis of combined β-thalassemia mutations and HLA typing. Of these, one embryo was diagnosed as homozygous normal for the thalassemia mutation and HLA matched with the existing affected sibling. Conclusion The optimized protocol worked well in PGD clinical cycle for selection of thalassemia-unaffected embryos with the desired HLA system.
ISSN:0188-4409
1873-5487
DOI:10.1016/j.arcmed.2016.07.006