Prevalence and molecular characterization of G6PD deficiency in two Plasmodium vivax endemic areas in Venezuela: predominance of the African A-202A/376G variant

Table 1 PCR/RFLP conditions used to identify the G6PD genetic variants of G6PDd unrelated individuals in the present study

Mutations identified by PCR/RFLP^a	Exon	Primer oligonucleotide sequence (5′–3′)	Amplicon size^b (bp)	References	RFLP pattern (fragment size in bp)
Mutations identified by PCR/RFLP^a	Exon	Primer oligonucleotide sequence (5′–3′)	Amplicon size^b (bp)	References	RE^c	Wild-type	Mutant^d
202G → A	4	GTGGCTGTTCCGGGATGGCCTTCTG CTTGAAGAAGGGCTCACTCTGTTTTG	109	29	NlaIII	109	63, 46
376A → G	5	CAGTACGATGATGCAGC CAGGTAGAAGAGGCGGT	90	29	FokI	90	58, 32
563C → T	6/7	ACTCCCCGAAGAGGGGT CCAGCCTCCCAGGAGAGA	542	30	MboII	25, 26, 119,377	25, 26, 100, 119, 277
844G → C	8	GGAGCTAAGGCGAGCTC CATGCTCTTGGGGACTG	230	30	NlaIII	11, 34, 75,110	11, 28, 47, 34,110

RFLP restriction fragment length polymorphism analysis, bp base pairs, RE restriction endonuclease used for RFLP
^aThe remaining mutations located in the exons 4–8, specifically the 185C → A, 542A → T, 592C → T, 593G → C, 634A → G, 637G → T, 680G → A, 820G → A, 835A → T, 854G → A and 871G → A, were studied by DNA sequencing
^bCycling conditions used in the PCR were as follows: 40 cycles at 94 °C for 1 min, 60 °C (for exon 4 and 8) or 56 °C (for exon 5) or 58 °C (for exon 6/7) for 30 s, and 72 °C for 40 s. A final elongation at 72 °C for 7 min was added
^cRestriction endonuclease digestion carried out with 5 U of enzyme at 37 °C for 3 h
^d NlaIII, FokI or MboII recognition site is created by the mutation

ISSN: 1475-2875