AIM: To survey the prevalence of toxoplasmosis in tumor patients in Hubei and to examine the significance of direct immunofixation agglutination (DIFA), IgG-IFAT, and ISAGA for dignosing Toxoplasma infection. METHODS: Sera from 100 tumor cases were tested by DIFA for detecting anti-Toxoplasma IgG antibodies. The positive sera were re-examined by IgG-IFAT and then detected by Toxo-IgG/IgA/IgM-ISAGA. RESULTS: Using DIFA, the total anti-Toxoplasma antibody positive rate was 12% (12/100). Using IgG-IFAT, the corresponding rate was 100%, with one case suspected to be IgA positive. Four cases were IgM positive, two of them were with active infection and one was dubious. The rates of Toxoplasma infection in patients with lung cancer, non-Hodgkin lymphoma and breast cancer were 22.2% (6/27), 19.3% (5/26) and 7.1% (1/14), respectively. CONCLUSION: DIFA is more sensitive and specific for diagnosing Toxoplasma infection and that cancer patients are more susceptible to Toxoplasma infection.