Background: Glucose 6-phosphate dehydrogenase deficiency (G6PDd) is an X-linked hereditaryrngenetic defect, affects an estimated 400 million people worldwide. Severe clinical manifestationrnassociated with G6PDd (e.g., chronic hemolytic anemia) depends on the type of G6PD molecularrnvariants and exposure to hemolytic triggers (e.g., antimalarial like Primaquine). However, scarcernstudies on G6PDd renders the use of Primaquine for effective therapeutic treatment of malaria.rnObjective: To determine the availability and characterize selected molecular variants of G6PDdrnspecific genes among selected populations in malaria endemic area of Ethiopia.rnMethod: A cross sectional study was conducted among selected populations in malaria endemicrnareas of Ethiopia from July 30, 2014 to January 30, 2015. A total of 523 dried blood spot samplesrnwere randomly selected from stored samples of national malaria indicator survey of 2011.rnPolymerase chain reaction and restricted fragment length polymorphism technique was applied torncharacterize G6PDd variants as G6PD*A, G6PD*A- and/or G6PD*Mediterranean. Binary logisticrnregression was applied to see association (P0.05).rnConclusion: G6PD*A variant was the only G6PDd genotype detected in this study. G6PD*Arnvariant has almost (90%) the same enzymatic activities with the wild type. Therefore; this resultrnsupports the safe use of primaquine, especially the single low dose for transmission interruptionrnof Plasmodium falciparum gametocyte and radical cure of Plasmodium vivax, as a part of malariarnelimination toolkit, among selected populations in malaria endemic areas of Ethiopia.rnKey terms: Glucose 6-phosphate Dehydrogenase deficiency, Polymerase Chain Reaction,rnRestriction Fragment Length Polymorphism, Malaria, Primaquine, Ethiopia