Background: - Glucose-6-phosphate dehydrogenase deficiency (G6PDd) is common in malariarnendemic regions that hinder the use of 8-aminoquinoline drugs as a radical cure for malaria duernto the risk of inducing hemolytic anemia.rnObjective: - To investigate G6PDd among apparently healthy individuals in selected malariarnendemic areas from different agroecological zones of Ethiopia.rnMethods: - a community based cross sectional survey involving 1609 individuals was donernusing genotypic and phenotypic analysis. CareStartTM Rapid Diagnostic kit (RDT) was used tornscreen for G6PD enzyme activity. Sequencing and polymerase chain reaction based restrictionrnfragment length polymorphism were done for further confirmation for all phenotypicallyrndetected enzymatic deficiencies and screened representative samples from those which testedrnphenotypically normal. Dried Blood Spot was collected for molecular analysis. Phenotypicallyrndeficient individuals were genotyped for the mutations, G202A, A376G and C563T. Plasmodiumrnblood-stage parasitaemia detection was performed using the CareStartTM Malaria AgrnPLDH/HRP2 and nested Polymerase Chain Reaction.rnResults: - G6PDd detected using the phenotypic approach was less (22/1609, 1.4%) than therngenotypic approach (31/222, 14%). Of the 22 G6PDd individuals detected by CareStartTM RDT,rn13 (1.50%) males and 9 (1.21%) females, 16/400 (4.00%), 4/484 (0.8%) and 2/401 (0.5%) werernfrom Gambella, Oromiya and Benishangule Gumuz respectively. Moreover, the G6PDdrnphenotypic prevalence was significantly higher 6.50 % (13/200) in the Agnuwak ethnic groupsrn(x2 =47.3431 and P = 0.001). Of the 31 individuals found to be G6PDd by sequencing, 29% (9)rnhemizygous males, 16.13% (5) homozygous females and 54.84% (17) heterozygous femalesrnwere found for 376A+ mutation. The highest asymptomatic malaria infection detected with RDTrnwas P. falciparum.rnConclusion: - This study found high Genetic diversity (14%) across the G6PD gene in the studyrnpopulation. As the use of currently available radical cures (gametocidal drugs) againstrnplasmodium are known to induce hemolytic anemia, further studies in larger groups needs to berndone. In this study the limited number (222/1609) of samples sequenced from all study sitesrnresulted in higher number of G6PDd individuals. .rnKeywords: - G6PD, G6PDd, Haemolytic anaemia, DNA sequencing, Haplotype, Gene mutation,rnMalaria, Ethiopia.