title: Complex preimplantation genetic diagnosis for beta-thalassaemia, sideroblastic anaemia, and human leukocyte antigen (HLA)-typing.
abstract: Preimplantation genetic diagnosis (PGD) to select histocompatible siblings to facilitate curative haematopoeitic stem-cell transplantation (HSCT) is now an acceptable option in the absence of an available human leukocyte antigen (HLA) compatible donor. We describe a case where the couple who requested HLA-PGD, were both carriers of two serious haematological diseases, beta-thalassaemia and sideroblastic anaemia. Their daughter, affected with sideroblastic anaemia, was programmed to have HSCT. A multiplex-fluorescent-touchdown-PCR protocol was optimized for the simultaneous amplification of: the two HBB-gene mutated regions (c.118C> T, c.25-26delAA), four short tandem repeats (STRs) in chr11p15.5 linked to the HBB gene, the SLC25A38 gene mutation (c.726C > T), two STRs in chr3p22.1 linked to the SLC25A38 gene, plus eleven informative STRs for HLA-haplotyping (chr6p22.1-21.3). This was followed by real-time nested PCR and high-resolution melting analysis (HRMA) for the detection of HBB and SLC25A38 gene mutations, as well as the analysis of all STRs on an automatic genetic analyzer (sequencer). The couple completed four clinical in vitro fertilization (IVF)/PGD cycles. At least one matched unaffected embryo was identified and transferred in each cycle. A twin pregnancy was established in the fourth PGD cycle and genotyping results at all loci were confirmed by prenatal diagnosis. Two healthy baby girls were delivered at week 38 of pregnancy. The need to exclude two familial disorders for HLA-PGD is rarely encountered. The methodological approach described here is fast, accurate, clinically-validated, and of relatively low cost.
authors.affiliation: a Department of Medical Genetics , National and Kapodistrian University of Athens, Choremio Research Laboratory .
authors.forename: Georgia
authors.initials: G
authors.lastname: Kakourou
authors.affiliation: a Department of Medical Genetics , National and Kapodistrian University of Athens, Choremio Research Laboratory .
authors.forename: Christina
authors.initials: C
authors.lastname: Vrettou
authors.affiliation: b First Department of Pediatrics , Athens University Medical School .
authors.forename: Antonis
authors.initials: A
authors.lastname: Kattamis
authors.affiliation: c Research Institute for the Study of Genetic and Malignant Disorders in Childhood, "Aghia Sophia" Children's Hospital .
authors.forename: Aspasia
authors.initials: A
authors.lastname: Destouni
authors.affiliation: a Department of Medical Genetics , National and Kapodistrian University of Athens, Choremio Research Laboratory .
authors.forename: Myrto
authors.initials: M
authors.lastname: Poulou
authors.affiliation: c Research Institute for the Study of Genetic and Malignant Disorders in Childhood, "Aghia Sophia" Children's Hospital .
authors.forename: Maria
authors.initials: M
authors.lastname: Moutafi
authors.affiliation: d Centre for Human Reproduction, Genesis Hospital , and.
authors.forename: Georgia
authors.initials: G
authors.lastname: Kokkali
authors.affiliation: d Centre for Human Reproduction, Genesis Hospital , and.
authors.forename: Konstantinos
authors.initials: K
authors.lastname: Pantos
authors.affiliation: e Embryogenesis, Centre for Human Reproduction , Athens , Greece.
authors.forename: Stephen
authors.initials: S
authors.lastname: Davies
authors.affiliation: a Department of Medical Genetics , National and Kapodistrian University of Athens, Choremio Research Laboratory .
authors.forename: Sophia
authors.initials: S
authors.lastname: Kitsiou-Tzeli
authors.affiliation: a Department of Medical Genetics , National and Kapodistrian University of Athens, Choremio Research Laboratory .|c Research Institute for the Study of Genetic and Malignant Disorders in Childhood, "Aghia Sophia" Children's Hospital .
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Kanavakis
authors.affiliation: a Department of Medical Genetics , National and Kapodistrian University of Athens, Choremio Research Laboratory .
authors.forename: Joanne
authors.initials: J
authors.lastname: Traeger-Synodinos
mesh_terms: D000328:Adult; D000756:Anemia, Sideroblastic / Q000175:diagnosis* / Q000235:genetics* / Q000628:therapy; D005260:Female; D005307:Fertilization in Vitro; D005820:Genetic Testing; D005838:Genotype; D018380:Hematopoietic Stem Cell Transplantation; D006650:Histocompatibility Testing / Q000379:methods*; D006801:Humans; D007231:Infant, Newborn; D008297:Male; D033681:Mitochondrial Membrane Transport Proteins / Q000096:biosynthesis / Q000235:genetics; D009154:Mutation / Q000235:genetics; D011247:Pregnancy; D059285:Pregnancy, Twin; D019836:Preimplantation Diagnosis / Q000379:methods*; D017086:beta-Thalassemia / Q000175:diagnosis* / Q000235:genetics* / Q000628:therapy
chemical_list: D033681:Mitochondrial Membrane Transport Proteins; C553226:Slc25a38 protein, human
keywords: Beta-thalassaemia; high resolution melting analysis; preimplantation genetic diagnosis; sideroblastic anaemia; single cell HLA-typing