title: Rapid detection of the major Mediterranean beta-thalassaemia mutations by real-time polymerase chain reaction using fluorophore-labelled hybridization probes.
abstract: We present a new method to detect the major beta-thalassaemia mutations of the Mediterranean countries (IVS I-1, IVS I-6, IVS I-110, CD-37 and CD-39). The procedure is based upon real-time polymerase chain reaction (PCR), using specific fluorescently labelled hybridization probes. The melting curves for each of the specific probes obtained after PCR enabled the identification of different alleles. Genotyping of 71 patients with thalassaemia and 20 controls without thalassaemia by the established method and conventional allele-specific restriction enzyme analysis produced identical results. The established method is a robust, fast and straightforward assay that allows detection of the major Mediterranean beta-thalassaemia mutations simultaneously in less than 60 min.
authors.affiliation: Laboratorio de Biología Molecular, Dpto. Biopatología Clínica, Hospital Universitario La Fe, Valencia, Spain.
authors.forename: Isabel
authors.initials: I
authors.lastname: Moreno
authors.forename: Pascual
authors.initials: P
authors.lastname: Bolufer
authors.forename: M Luz
authors.initials: ML
authors.lastname: Perez
authors.forename: Eva
authors.initials: E
authors.lastname: Barragán
authors.forename: Miguel A
authors.initials: MA
authors.lastname: Sanz
mesh_terms: D000483:Alleles; D001483:Base Sequence; D016022:Case-Control Studies; D004252:DNA Mutational Analysis; D015342:DNA Probes / Q000235:genetics; D005456:Fluorescent Dyes; D005838:Genotype; D005914:Globins / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity; D017086:beta-Thalassemia / Q000235:genetics*
chemical_list: D015342:DNA Probes; D005456:Fluorescent Dyes; D005914:Globins