title: Novel one-step multiplex PCR-based method for HLA typing and preimplantational genetic diagnosis of β-Thalassemia.
abstract: Preimplantation genetic diagnosis (PGD) of single gene disorders, combined with HLA matching (PGD-HLA), has emerged as a tool for couples at risk of transmitting a genetic disease to select unaffected embryos of an HLA tissue type compatible with that of an existing affected child. Here, we present a novel one-step multiplex PCR to genotype a spectrum of STRs to simultaneously perform HLA typing and PGD for β-thalassemia. This method is being routinely used for PGD-HLA cycles in our department, with a genotyping success rate of 100%. As an example, we present the first successful PGD-HLA typing in Spain, which resulted in the birth of a boy and subsequent successful HSC transplantation to his affected brother, who is doing well 4 years following transplantation. The advantage of our method is that it involves only a round of single PCR for multiple markers amplification (up to 10 markers within the HLA and 6 markers at the β-globin loci). This strategy has allowed us to considerably reduce the optimization of the PCR method for each specific PGD-HLA family as well as the time to obtain molecular results in each cycle.
authors.affiliation: Department of Genetics, Reproduction and Fetal Medicine, Institute of Biomedicine of Seville (IBIS), University Hospital Virgen del Rocío/CSIC/University of Seville, Avenida Manuel Siurot, s/n, 41013 Seville, Spain.
authors.forename: Raquel M
authors.initials: RM
authors.lastname: Fernández
authors.forename: Ana
authors.initials: A
authors.lastname: Peciña
authors.forename: Maria Dolores
authors.initials: MD
authors.lastname: Lozano-Arana
authors.forename: Juan Carlos
authors.initials: JC
authors.lastname: García-Lozano
authors.forename: Salud
authors.initials: S
authors.lastname: Borrego
authors.forename: Guillermo
authors.initials: G
authors.lastname: Antiñolo
mesh_terms: D000328:Adult; D002648:Child; D004624:Embryo Transfer; D005260:Female; D005307:Fertilization in Vitro; D006650:Histocompatibility Testing / Q000379:methods*; D006801:Humans; D008297:Male; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D009865:Oocytes / Q000166:cytology / Q000378:metabolism; D011247:Pregnancy; D019836:Preimplantation Diagnosis; D027724:Reproductive Techniques, Assisted; D013030:Spain; D055544:beta-Globins / Q000235:genetics*; D017086:beta-Thalassemia / Q000097:blood / Q000175:diagnosis* / Q000235:genetics
chemical_list: D055544:beta-Globins