Background. Toxoplasma gondii is an obligate intracellular protozoan parasite and is a major opportunistic pathogen in immune-compromised hosts. This study assessed the prevalence of T. gondii and associated risk factors among people living with HIV. Methods and Materials. A cross-sectional study was carried out among people living with HIV attending Gondar University Hospital. A structured and pretested questionnaire was used to collect sociodemographic factors, and 10mL of venous blood was collected for anti-Toxoplasma antibody test and determination of CD4 levels. Serum was tested in duplicate for anti-Toxoplasma antibody using rapid slide agglutination test. Results. A total of 170 study subjects were enrolled in the study. Seroprevalence of T. gondii among the study participants was 76.5% (95% CI: 69.0–82.8). High proportions of seropositive individuals (64.7%) were found under the child bearing age groups. The mean CD4+ lymphocyte count of HIV monoinfected participants was cells/mm3 while coinfected study participants had mean CD4+ lymphocyte count of cells/mm3 with value of 0.01. Conclusion. The seroprevalence of T. gondii among people living with HIV was high. Cautious followup of HIV-positive patients is needed to prevent development of toxoplasmic encephalitis and other related complications.