OBJECTIVE: The enzyme glucose-6-phosphate dehydrogenase (G6PD) catalyses the 
first step in the pentose phosphate pathway, producing nicotinamide adenine 
dinucleotide phosphate (NADPH). NADPH plays a crucial role in preventing 
oxidative damage to proteins and other molecules in cells, mostly red blood 
cells. G6PD deficiency has an x-linked pattern of inheritance in which 
hemizygous males are deficient, while females may or may not be deficient 
depending on the number of affected alleles. We report two novel DNA variants in 
the G6PD gene detected in two male probands with chronic nonspherocytic 
hemolytic anemia (CNSHA), who were referred for hematological evaluation.
METHOD: Probands and their relatives underwent clinical, biochemical, and 
molecular assessment.
RESULTS: Two novel DNA variants, c.995C>T and c.1226C>A, were found in this 
study. At the protein level, they produce the substitution of Ser332Phe and 
Pro409Gln, respectively. These DNA variants were analyzed in the female 
relatives of probands for genetic counseling.
CONCLUSIONS: The novel DNA variants were classified as class I based on the 
clinical, biochemical, and molecular evaluations performed.