title: Application of MLPA assay to characterize unsolved α-globin gene rearrangements.
abstract: α-thalassemia belongs to those inherited diseases in which large genomic deletions/duplications represent a significant proportion of causative mutations. Until recently, large α-globin gene cluster rearrangements have been mainly detected by gap-PCR and Southern blotting, methods that have significant drawbacks. We tested the recently developed multiplex ligation-dependent probe amplification (MLPA) assay for deletional screening of the α-globin gene cluster in a cohort of 25 individuals suspected of having α-globin alteration(s), in which no or doubtful mutations had been found using conventional methods. In 13 out of 18 α-thalassemia carriers and in all 5 patients with HbH we found the causative α-globin defects. In 2 thalassemia intermedia patients, carriers of heterozygous β-globin mutations, the co-inheritance of homozygous α-genes triplication was detected. MLPA results were subsequently confirmed by real-time PCR. This study shows that MLPA can effectively identify different and unknown types of α-globin gene rearrangements, to allow characterizing previously unsolved α-thalassemia genotypes.
authors.affiliation: Dept. of Comparative Biomedical Sciences, University of Teramo, Italy. acolosimo@unite.it
authors.forename: Alessia
authors.initials: A
authors.lastname: Colosimo
authors.forename: Valentina
authors.initials: V
authors.lastname: Gatta
authors.forename: Valentina
authors.initials: V
authors.lastname: Guida
authors.forename: Eleonora
authors.initials: E
authors.lastname: Leodori
authors.forename: Enrica
authors.initials: E
authors.lastname: Foglietta
authors.forename: Silvana
authors.initials: S
authors.lastname: Rinaldi
authors.forename: Maria Pia
authors.initials: MP
authors.lastname: Cappabianca
authors.forename: Antonio
authors.initials: A
authors.lastname: Amato
authors.forename: Liborio
authors.initials: L
authors.lastname: Stuppia
authors.forename: Bruno
authors.initials: B
authors.lastname: Dallapiccola
mesh_terms: D000328:Adult; D000368:Aged; D001681:Biological Assay; D015139:Blotting, Southern; D002675:Child, Preschool; D005260:Female; D018628:Gene Dosage; D015321:Gene Rearrangement; D056726:Genetic Association Studies; D005838:Genotype; D006579:Heterozygote; D006801:Humans; D022521:Ligase Chain Reaction / Q000379:methods; D008297:Male; D005810:Multigene Family; D009154:Mutation; D010641:Phenotype; D016133:Polymerase Chain Reaction; D055815:Young Adult; D055542:alpha-Globins / Q000235:genetics* / Q000378:metabolism; D017085:alpha-Thalassemia / Q000175:diagnosis / Q000235:genetics*
chemical_list: D055542:alpha-Globins