MOLECULAR ANALYSIS FOR THE DETECTION OF GLUCOSE 6- PHOSPHATE DEHYDROGENASE (G6PD) DEFICIENCY

ABSTRACT Glucose-6-Phosphate Dehydrogenase is the first and primary enzyme of the regulatory enzymes in the pentose phosphate pathway - the only method by which red blood cells can generate NADPH. Deficiency of G6PD is the most common enzymopathy that affects over 400 million people worldwide. Glucose-6-Phosphate Dehydrogenase deficiency is found in regions where malaria is endemic or has been endemic and it confers resistance against malaria especially malaria caused by Plasmodium falciparum. Mutation upon this X-linked gene - encoding for G6PD - causes enzyme deficiency from asymptomatic to chronic nonspherocytic haemolytic anemia. Over 300 biochemical and genetically distinct variants of G6PD deficiency have been described in the past due to the polymorphism of the gene. This study intends to assess the molecular analysis for the detection of G6PD deficiency in given areas namely Medan, Bangka Island and Palangkaraya, where the level of malaria is not very high but G6PD deficiency is present. Blood samples were collected from apparently healthy males. Glucose-6-Phosphate Dehydrogenase deficiency was screened biochemically by using Sigma fluorescence spot test. DNA was extracted from buffycoat using soluble saturated phenol (ss-phenol) standard method while exons were PCR- amplified. Restriction endonudease digestion analysis was used to determine known mutations while Single-Strand Conformation Polymorphism (SSCP) followed by DNA sequencing was used to determine unknown mutations. DNA from deficient males was the subject for this study and DNA from normal male was used as control for all analysis. From 13 deficient male subjects studied, subjects from Bangka Island and Palankaraya showed G6PD deficiency on molecular level. Glucose-6¬Phosphate Dehydrogenase Canton variant was found in Bangka. The mutation produces a Gâ�