Abstract: Toxoplasma gondii is one of the most common parasites that cause infection to human. The aim of this study was to conduct a sero-prevalence survey of toxoplasmosis in several target groups. A total of 1146 serum samples were collected and detected against Toxoplasma gondii antibodies using LAT. Five hundred (out of 1146 cases) were positive cases giving prevalence rate of (43.6%). High a prevalence was found among HIV patients (75%), aborters (58.3%) and suspected cases (55.5%). More positive cases were detected by (LAT) in age groups (26-30) years than other groups (p = 0.0001). Prevalence rate by sex was found to be higher in females than males (p<0.05). Residence was found to be significant in rural areas (p = 0.001). The relationship between positive (LAT) and women who aborted was significant (p<0.05). Contact with cats, eating raw meat and eating soil were significant to infection using (LAT) (p = 0.0001, p = 0.0001 and p = 0.006), respectively. High prevalence of Toxoplasma gondii was recorded in Khartoum State.