Rapid genotyping of two common G6PD variants, African (A-) and Mediterranean, by high-resolution melting analysis.
OBJECTIVE
The 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.


METHODS
After 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.


RESULTS
Wild, 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.


CONCLUSIONS
Our HRM protocols allow a rapid, simple and cost-effective screening of G6PD deficiency in patients originating from the Mediterranean and the African areas.
Joly
Philippe
P
Unité de Pathologie Moléculaire du Globule Rouge, Fédération de Biochimie et de Biologie Spécialisée, Hôpital Edouard Herriot, Hospices Civils and Université Claude Bernard-Lyon 1, Lyon, France.
Lacan
Philippe
P
Garcia
Caroline
C
Martin
Cyril
C
Francina
Alain
A
D000349:Africa; D005060:Europe; D005260:Female; D005820:Genetic Testing / Q000191:economics / Q000379:methods; D014644:Genetic Variation; D005838:Genotype; D005954:Glucosephosphate Dehydrogenase / Q000235:genetics*; D005955:Glucosephosphate Dehydrogenase Deficiency / Q000235:genetics*; D006801:Humans; D008297:Male; D009154:Mutation; D009691:Nucleic Acid Denaturation; D044469:Racial Groups / Q000235:genetics
D005954:Glucosephosphate Dehydrogenase