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Background: Toxoplasma gondii is one of the most prevalent zoonotic opportunistic parasitic infections in the world. The aim of this study was to examine chronic and acute toxoplasmosis molecularly and serologically in Hemodialysis patients.Methods: A total of 238 serum samples of hemodialysis patients with chronic renal disease (119 samples) as case group and healthy individuals (119 samples) as control group were enrolled in Zahedan City, southeastern Iran. The molecular and serological detection of toxoplasmosis were conducted on all samples using nested-PCR and ELISA.Results: The prevalence rates of anti-T. gondii IgM and IgG and T. gondii DNA in the case group were respectively 0%, 44.5% and 29.4% and the corresponding values in the control group were 0.8%,23.5% and 2.52 % respectively (p≥0.05).Conclusion: In conclusion, preliminary Toxoplasma gondii infection screening is required using serological techniques, particularly in hemodialysis patients who are frequently exposed to hemodialysis so as to stop infection dissemination.