Rapid and inexpensive detection of common HBB gene mutations in Tunisian population by high-resolution melting analysis: Implication for molecular diagnosis |
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Authors: | Houyem Ouragini Faten Haddad Imen Darragi Salem Abbes |
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Affiliation: | Laboratoire d'Hématologie Moléculaire et Cellulaire (LR O5 SP 04), Institut Pasteur de Tunis, Tunis 1002, Tunisie |
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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. |
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Keywords: | β-Thalassemia HBB gene Common mutations High-resolution melting Mutation analysis Molecular diagnosis |
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