Molecular genetics of glucose‐6‐phosphate dehydrogenase (G6PD) deficiency in Spain: identification of two new point mutations in the G6PD gene

Abstract

In order to explore the nature of glucose-6-phosphate dehydrogenase (G6PD) deficiency in Spain, we have analysed the G6PD gene in 11 unrelated Spanish G6PD-deficient males and their relatives by using the polymerase chain reaction and single-strand conformation polymorphism (PCR-SSCP) analysis combined with a direct PCR-sequencing procedure and PCR-restriction enzyme (RE) analysis. We have identified eight different missense mutations, six of which have been reported in previously described G6PD variants. In nine patients who had presented with acute favism we found the following mutations: G6PD A-376G-202A (four cases), G6PD Union1360T (two cases), G6PD Mediterranean563T (one case) and G6PD Aures143C (one case). In the remaining patient a novel A to G transition was found at nucleotide position 209 which has not been reported in any other ethnic group. This mutation results in a (70) Tyr to Cys substitution and the resulting G6PD variant was biochemically characterized and designated as G6PD Murcia. This new mutation creates a Bsp 1286I recognition site which enabled us to rapidly detect it by PCR-RE analysis. In two patients with chronic non-spherocytic haemolytic anaemia (CNSHA) we found the underlying genetic defects, as had been noted previously, to be located within a cluster of mutations in exon 10. One of them had the T to C transition at nucleotide 1153, causing a (385) Cys to Arg substitution, previously described in G6PD Tomah. The other, previously reported as having a variant called G6PD Clinic, has a G to A transition at nucleotide 1215 that produces a (405) Met to Ile substitution, thus confirming that G6PD Clinic is a new class I variant.

Molecular characterization of G6PD deficiency in Southern Italy: heterogeneity, correlation genotype–phenotype and description of a new variant (G6PD Neapolis)

“…Our study clearly demonstrates that the mutation 188 Ser →Phe (G6PD Mediterranean) is strongly associated with haemolytic events whereas the mutation 282 Asp →Hys (Seattle) is always associated with a non-haemolytic phenotype (Table I). This is in agreement with a study of nine unrelated Spanish subjects with favism, none of whom was found to bear the G6PD Seattle variant, suggesting that this variant, which is polymorphic in Spain, is asymptomatic (Rovira et al, 1995). As for other variants, the small number of cases does not allow us to draw meaningful conclusions regarding the haemolytic phenotype.…”

Glucose-6-phosphate Dehydrogenase Deficiency in Portugal: Biochemical and Mutational Profiles, Heterogeneity, and Haplotype Association

“…Recently, by the use of molecular analysis, the G6PD Seattle mutation was found in Spain (28), in the Canary Islands (29,30) and in the north African Berber (30). Nafa et al (13) found the Seattle mutation associated with haplotype (Ϫ Ϫ ϩ ϩ Ϫ Ϫ) in Algeria and, and Medina et al found in England an association to the same haplotype although restricted to four RFLPs (PvuII, PstI, BclI, and NlaIII) (31).…”

Independent origin of single and double mutations in the human glucose 6-phosphate dehydrogenase gene

“…Among these, G6PD Mediterranean (563 C-+T), G6PD A-(376 A+G with either 202 G+A or 968 T-C), G6PD Seattle (844 G+C), G6PD Aures (143 T-C), and G6PD Union (1360 C+T), have been found in Spain (Rovira et al, 1994(Rovira et al, , 1995. Here, the G6PD gene has been characterized in five unrelated Spanish G6PD deficient males who had presented clinically with favism.…”

“…The DNA region from the G6PD gene encompassing each point mutation was selectively amplified by PCR as described previously (Poggi et al, 1990;Rovira et al, 1995). Naturally occurring restriction sites enabled us to distinguish the normal from mutant genes as follows: Fok I for the 376 A+G mutation of G6PD A and G6PD A-; Nla 111 for the 202 G-+A mutation of the most common form of G6PD A-; Bst NI for the 680 G+T mutation and Nci I for the 968 T+C mutation of the two additional G6PD A-genotypes; Mbo I1 for the 563 C+T mutation of G6PD Mediterranean; and Hha I for the 1360 C+T mutation of G6PD Union.…”