Background: Toxoplasmosis is a universal opportunistic infection that causes severe complications in immunosuppressed patients. Renal failure is a state of immunodeficiency, especially in children. Therefore, hemodialysis patients may be at high risk for toxoplasmosis. Objective: To investigate the seropositivity rate of T. gondii specific antibodies (anti-T. gondii IgM and IgG antibodies) in pediatric hemodialysis patients. Subjects and Methods: ELISA was used to test serum samples from 67 children on regular hemodialysis and 50 healthy controls for anti-Toxoplasma IgG and IgM antibodies in this case-control research. Demographic criteria, duration of hemodialysis, and possible risk factors for toxoplasmosis were recorded. All participants were clinically examined to detect any signs suggestive of toxoplasmosis. Results: In comparison to total T. gondii seropositivity in all participants (23%), it was 16% and 28% in control and hemodialysis groups, respectively. The IgG antibodies were detected in 19 hemodialysis cases and eight control cases. The mean duration of hemodialysis among T. gondii seropositive and seronegative patients were 47.6±14.3 and 22.3±6.7 months, respectively. Among T. gondii seropositive individuals, the average duration of hemodialysis was substantially longer. In the examined groups, contact with cats and eating semi-cooked meat were the risk factors recorded for T. gondii seropositivity. Conclusion: Toxoplasmosis seropositivity rate was higher in hemodialysis children than in the control group. For early identification and treatment, T. gondii screening should be done for all hemodialysis patients before and during hemodialysis.