Malaria is one of the most common diseases in the African population. Genetic variance in glucose dehydrogenase 6-phosphate (G6PD) in humans determines the response to malaria exposure. In this study, we aimed to analyze the frequency of two single-nucleotide polymorphisms (G202A and A376G) present in two local tribes of Sudanese Arabs from the region of the 4th Nile cataract in Sudan, the Shagia and Manasir. The polymorphisms in G6PD were analyzed in 217 individuals (126 representatives of the Shagia tribe and 91 of the Manasir tribe). Real-time PCR and RFLP-PCR were utilized to analyze significant differences in the prevalence of alleles and genotypes. The 202A G6P allele frequency was 0.7%, whereas the G202 variant was found in 93.3% of cases. The AA, GA, and GG genotype frequencies for the A376G G6PD codon among the Shagia were 88, 11.1, and 0.9%, respectively; this is similar to the distribution among Manasir tribe representatives (94.5, 3.3, and 2.2%, respectively; OR 3.44 [0.85-16.17], p=0.6). Notably, in north-eastern Sudan the G6PD B (202G/376A) compound genotype frequency was 90.3%, whereas the G6PD A variant (202G/376G) was found in 1.4% of that population. Identification of the G6PD A- variant (202A/376G) in the isolated Shagia tribe provides important information regarding the tribal ancestry. Taken together, the data presented in this study suggest that the Shagia tribe was still nomadic between 4000 and 12,000years ago. Moreover, the lack of G6PD A- genotype among ethnically diverse Monasir tribesmen indicates a separation of the Shagia from the other tribes in the region of the 4th Nile cataract in Sudan.