Background: Enuresis is one of the most common diseases in children. Although there are several factors involved in the occurrence of this diseases, the root cause of it has remained undetermined. Objectives: Identifying various factors responsible for enuresis may enormously contribute to solving this problem. This study, therefore, aimed to determine the level of vitamin B12 and folic acid in children with enuresis in Gorgan in 2021. Methods: In this case-control study, 43 children with primary enuresis and 99 children without enuresis, as the control group, referring to Taleghani Hospital in 2021 were included. Folic acid and vitamin B12 levels in two groups were measured and analyzed using statistical techniques. Results: Out of all participants, 23 (53.5%) in the case group and 53 (53.5%) in the control group were male. The mean age of children in the case and control groups were 7.60 ± 3.02 and 8.93 ± 3.15, respectively, and the two groups were not significantly different in terms of gender and age. Vitamin B12 and folic acid levels in the case group were significantly lower than those in the control group (P-value = 0.001). There was a significant difference between the case and control groups regarding the mean levels of vitamin B12 and folic acid levels based on sex and gender. Conclusions: In sum, it was found that children with enuresis suffered from deficiency of vitamin B12 and folic acid to some extent, which may have been a factor responsible for delaying the maturation of the central nervous system and, consequently, inducing enuresis in children.