title: Development of a High-Resolution Melting Approach for Scanning Beta Globin Gene Point Mutations in the Greek and Other Mediterranean Populations.
abstract: Beta-thalassaemia is one of the most common autosomal recessive disorders worldwide. The disease's high incidence, which is observed in the broader Mediterranean area has led to the establishment of molecular diagnostics' assays to prevent affected births. Therefore, the development of a reliable, cost-effective and rapid scanning method for β globin gene point mutations, easily adapted to a routine laboratory, is absolutely essential. Here, we describe, for the first time, the development of a High-Resolution Melting Analysis (HRMA) approach, suitable for scanning the particularly heterogeneous beta globin gene mutations present in the Greek population, and thus adaptable to the Mediterranean and other areas where these mutations have been identified. Within this context, β globin gene regions containing mutations frequently identified in the Greek population were divided in ten overlapping amplicons. Our reactions' setup allowed for the simultaneous amplification of multiple primer sets and partial multiplexing, thereby resulting in significant reduction of the experimental time. DNA samples from β-thalassaemia patients/carriers with defined genotypes were tested. Distinct genotypes displayed distinguishable melting curves, enabling accurate detection of mutations. The described HRMA can be adapted to a high-throughput level. It represents a rapid, simple, cost-effective, reliable, highly feasible and sensitive method for β-thalassaemia gene scanning.
authors.affiliation: Molecular Genetics Laboratory, Thalassaemia and Hemoglobinopathies Center, Laiko General Hospital, Athens, Greece.
authors.forename: Christos
authors.initials: C
authors.lastname: Chassanidis
authors.affiliation: Molecular Genetics Laboratory, Thalassaemia and Hemoglobinopathies Center, Laiko General Hospital, Athens, Greece.
authors.forename: Effrossyni
authors.initials: E
authors.lastname: Boutou
authors.affiliation: Thalassaemia and Hemoglobinopathies Center, Laiko General Hospital, Athens, Greece.
authors.forename: Ersi
authors.initials: E
authors.lastname: Voskaridou
authors.affiliation: Molecular Genetics Laboratory, Thalassaemia and Hemoglobinopathies Center, Laiko General Hospital, Athens, Greece.
authors.forename: Angeliki
authors.initials: A
authors.lastname: Balassopoulou
mesh_terms: D016022:Case-Control Studies; D005820:Genetic Testing / Q000379:methods*; D005838:Genotype; D006115:Greece; D006579:Heterozygote; D006801:Humans; D009691:Nucleic Acid Denaturation; D017354:Point Mutation; D012680:Sensitivity and Specificity; D055544:beta-Globins / Q000235:genetics*; D017086:beta-Thalassemia / Q000175:diagnosis* / Q000235:genetics
chemical_list: D055544:beta-Globins