title: Rapid and inexpensive detection of common HBB gene mutations in Tunisian population by high-resolution melting analysis: implication for molecular diagnosis.
abstract: In Tunisia, β-thalassemia is a common hereditary disease with a carrying rate of 2.21%. Up to now, detection of responsible mutations was made by laborious, expensive, and/or time consuming methods. The aim of this study is to develop and validate a specific assay for detection of the two most frequent mutations in Tunisian population, the IVS-I-110 (G → A) and Cd39 (C → T) mutations. In this study, we optimize high resolution melting analysis (HRMA) conditions for these mutations, using control DNAs. Then, we evaluate the strength of this methodology by screening a cohort of patients with β-thalassemia. All examined reference DNA samples were unambiguously distinguished from each other. For the blinded test, the results were completely compatible with direct sequencing, performed after the HRMA. As HRMA represents a highly sensitive and high-throughput gene scanning method, it can provide timely diagnosis at low cost for effective clinical management of β-thalassemia.
authors.forename: Houyem
authors.initials: H
authors.lastname: Ouragini
authors.forename: Faten
authors.initials: F
authors.lastname: Haddad
authors.forename: Imen
authors.initials: I
authors.lastname: Darragi
authors.forename: Salem
authors.initials: S
authors.lastname: Abbes
mesh_terms: D015331:Cohort Studies; D004247:DNA / Q000097:blood / Q000235:genetics*; D005787:Gene Frequency; D006454:Hemoglobins / Q000235:genetics*; D006801:Humans; D009154:Mutation; D057089:Pathology, Molecular; D017422:Sequence Analysis, DNA; D044366:Transition Temperature; D014416:Tunisia; D017086:beta-Thalassemia / Q000097:blood / Q000235:genetics*
chemical_list: D006454:Hemoglobins; D004247:DNA; C020622:hemoglobin B
keywords: Common mutations; HBB gene; High-resolution melting; Molecular diagnosis; Mutation analysis; β-Thalassemia