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Rapid detection of the major Mediterranean β‐thalassaemia mutations by real‐time polymerase chain reaction using fluorophore‐labelled hybridization probes
Author(s) -
Moreno Isabel,
Bolufer Pascual,
Perez M. Luz,
Barragán Eva,
Sanz Miguel A.
Publication year - 2002
Publication title -
british journal of haematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.907
H-Index - 186
eISSN - 1365-2141
pISSN - 0007-1048
DOI - 10.1046/j.1365-2141.2002.03823.x
Subject(s) - genotyping , polymerase chain reaction , microbiology and biotechnology , biology , real time polymerase chain reaction , allele , restriction enzyme , genotype , genetics , gene
Summary. We present a new method to detect the major β‐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 β‐thalassaemia mutations simultaneously in less than 60 min.