Rapid genotyping of two common G6PD variants,African (A-) and Mediterranean,by high-resolution melting analysis |
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| Authors: | Philippe Joly Philippe Lacan Caroline Garcia Cyril Martin Alain Francina |
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| Affiliation: | 1. Department of Pathology, Microbiology and Immunology, Vanderbilt University, Nashville, TN, United States;2. Department of Orthopaedic Surgery, David Geffen School of Medicine at UCLA, Los Angeles, CA, United States;3. School of Pharmacy, Centre for Biomolecular Sciences, University of Nottingham, Nottingham, UK;1. Allergy and Clinical immunology Unit, Hadassah-Hebrew University Medical Center, Jerusalem, Israel;2. Internal Medicine Division, Hadassah-Hebrew University Medical Center, Ein Kerem, Jerusalem, Israel;3. Pediatrics Department A, Soroka University Medical Center and Faculty of Health Sciences, Ben-Gurion University of the Negev, Beer Sheva, Israel;4. The Lautenberg Center for Immunology and Cancer Research, Institute of Medical Research Israel-Canada, Hebrew University-Hadassah Medical School, Jerusalem, Israel;5. Department of Pediatrics B, Schneider Children''s Medical Center of Israel, Petah Tiqva, Israel;6. Institute of Gastroenterology and Liver Diseases, Department of Medicine, Hadassah-Hebrew University Medical Center, Jerusalem, Israel;7. Department of Human Genetics and Metabolic Diseases, Hadassah-Hebrew University Medical Center, Jerusalem, Israel;8. Department of Clinical Biochemistry and Pharmacology, Faculty of Health Sciences, Ben-Gurion University of the Negev, Beer Sheva, Israel |
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| Abstract: | ObjectivesThe Mediterranean and A(-) G6PD variants are particularly prevalent in Africa and Southern Europe. Our study was aimed to develop an assay for the rapid genotyping of these two variants by HRM.MethodsAfter PCR reactions corresponding to the G6PD Mediterranean (exon 6), G6PD (A-) (exon 4) and G6PD (A-) (exon 5) mutations, amplicons were submitted to HRM. This protocol was applied to a cohort of 132 patients suffering from sickle cell disease.ResultsWild, homozygous or hemizygous and heterozygous states were fully discriminated by HRM for all three mutations. HRM results were in total accordance with DNA sequencing for 22 patients of our cohort with a ‘A’ genotype: presence of the (A-) (exon 5) mutation but absence of the (A-) (exon 4) mutation.ConclusionsOur HRM protocols allow a rapid, simple and cost-effective screening of G6PD deficiency in patients originating from the Mediterranean and the African areas. |
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