title: Setup of a Protocol of Molecular Diagnosis of β-Thalassemia Mutations in Tunisia using Denaturing High-Performance Liquid Chromatography (DHPLC).
abstract: BACKGROUND
  β-Thalassemia is one of the most prevalent worldwide autosomal recessive disorders. It presents a great molecular heterogeneity resulting from more than 200 causative mutations in the β-globin gene. In Tunisia, β-thalassemia represents the most prevalent monogenic hemoglobin disorder with 2.21% of carriers. Efficient and reliable mutation-screening methods are essential in order to establish appropriate prevention programs for at risk couples. The aim of the present study is to develop an efficient method based on the denaturing high-performance liquid chromatography (DHPLC) in which the whole β-globin gene (HBB) is screened for mutations covering about 90% of the spectrum.
  
  
  METHODS
  We have performed the validation of a DHPLC assay for direct genotyping of 11 known β-thalassemia mutations in the Tunisian population.
  
  
  RESULTS
  DHPLC assay was established based on the analysis of 62 archival β-thalassemia samples previously genotyped then validated with full concordance on 50 tests with blind randomized samples previously genotyped with DNA sequencing and with 96% of consistency on 40 samples as a prospective study.
  
  
  CONCLUSIONS
  Compared to other genotyping techniques, the DHPLC method can meet the requirements of direct genotyping of known β-thalassemia mutations in Tunisia and to be applied as a powerful tool for the genetic screening of prenatal and postnatal individuals.
authors.affiliation: Laboratory of Clinical Biochemistry and Molecular Biology, Research Laboratory LR00SP03, Children's Hospital, Tunis, Tunisia.
authors.forename: Chaima Abdelhafidh
authors.initials: CA
authors.lastname: Sahli
authors.affiliation: Laboratory of Clinical Biochemistry and Molecular Biology, Research Laboratory LR00SP03, Children's Hospital, Tunis, Tunisia.
authors.forename: Ikbel
authors.initials: I
authors.lastname: Ben Salem
authors.affiliation: Laboratory of Clinical Biochemistry and Molecular Biology, Research Laboratory LR00SP03, Children's Hospital, Tunis, Tunisia.
authors.forename: Latifa
authors.initials: L
authors.lastname: Jouini
authors.affiliation: Laboratory of Clinical Biochemistry and Molecular Biology, Research Laboratory LR00SP03, Children's Hospital, Tunis, Tunisia.
authors.forename: Naouel
authors.initials: N
authors.lastname: Laouini
authors.affiliation: Laboratory of Clinical Biochemistry and Molecular Biology, Research Laboratory LR00SP03, Children's Hospital, Tunis, Tunisia.
authors.forename: Rym
authors.initials: R
authors.lastname: Dabboubi
authors.affiliation: Laboratory of Clinical Biochemistry and Molecular Biology, Research Laboratory LR00SP03, Children's Hospital, Tunis, Tunisia.
authors.forename: Sondes
authors.initials: S
authors.lastname: Hadj Fredj
authors.affiliation: Laboratory of Clinical Biochemistry and Molecular Biology, Research Laboratory LR00SP03, Children's Hospital, Tunis, Tunisia.
authors.forename: Hajer
authors.initials: H
authors.lastname: Siala
authors.affiliation: Laboratory of Clinical Biochemistry and Molecular Biology, Research Laboratory LR00SP03, Children's Hospital, Tunis, Tunisia.
authors.forename: Rym
authors.initials: R
authors.lastname: Othmeni
authors.affiliation: Laboratory of Clinical Biochemistry and Molecular Biology, Research Laboratory LR00SP03, Children's Hospital, Tunis, Tunisia.
authors.forename: Boutheina
authors.initials: B
authors.lastname: Dakhlaoui
authors.affiliation: Laboratory of Clinical Biochemistry and Molecular Biology, Research Laboratory LR00SP03, Children's Hospital, Tunis, Tunisia.
authors.forename: Slaheddine
authors.initials: S
authors.lastname: Fattoum
authors.affiliation: Laboratory of Clinical Biochemistry and Molecular Biology, Research Laboratory LR00SP03, Children's Hospital, Tunis, Tunisia. amina.bibi@rns.tn.
authors.forename: Amina
authors.initials: A
authors.lastname: Bibi
authors.affiliation: Laboratory of Clinical Biochemistry and Molecular Biology, Research Laboratory LR00SP03, Children's Hospital, Tunis, Tunisia.
authors.forename: Taieb
authors.initials: T
authors.lastname: Messaoud
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004252:DNA Mutational Analysis; D005260:Female; D005820:Genetic Testing / Q000379:methods*; D005838:Genotype; D006801:Humans; D008297:Male; D009154:Mutation / Q000235:genetics*; D014416:Tunisia / Q000453:epidemiology; D055544:beta-Globins / Q000235:genetics*; D017086:beta-Thalassemia / Q000175:diagnosis* / Q000235:genetics
chemical_list: D055544:beta-Globins
keywords: DHPLC; mutation analysis; β-globin gene; β-thalassemia