Glucose phosphate dehydrogenasis (G6PD), the most prevalent enzymatic disease in humans, exists in south-eastern Iran. The geographic correlation of its distribution with the historic malaria endemic suggests that G6PD has increased in frequency as a result of natural selection by malaria. Based on studies, there is a controversy in terms of different analytical methods in terms of resistance to malaria. Fifty malaria patients and 50 healthy individuals from several cities south-east of Iran were included in the study and after obtaining consent, blood samples were taken from them. G6PD enzyme deficiency was investigated using a fluorescent stain test. The age, gender, and nationality of malaria patients were also assessed. The results were analyzed using SPSS software and appropriate statistical tests, and the value of P<0.05 was considered significant. The results showed that in malaria patients only one person had G6PD deficiency, while this number was six in the control group, which is significantly higher than the malaria group (P<0.05). Age group 27-42 years, men and people with Iranian citizenship also showed the highest incidence of malaria. Based on the results, it can be concluded that G6PD enzyme deficiency causes resistance to malaria and the frequency of this enzyme deficiency in malaria patients is significantly lower than in other people.