Toxoplasma gondii represents the most prominent infectious parasitic organism found in humans. While normally asymptomatic in healthy individuals, toxoplasmosis can cause abortion in patients during pregnancy, or can be fatal in immunosupressed individuals such as persons suffering from acquired immunodeficiency syndrom (AIDS). Toxoplasma gondii infection in humans is routinely assesssed by serological means. Here, we show that detection of anti-T. gondii IgG is also possible using a non-invasive methodology employing saliva. Sera and saliva of 201 healthy volunteers were investigated for the presence of anti-T. gondii-IgG antibodies by immunoblotting. The sera of 59 (29.4%) individuals showed IgG antibodies against T. gondii by ELISA, Vidas, and immunoblotting; 58 (98.3%) of these were also positive for anti-T. gondii IgG in the saliva immunoblot, with diagnostic relevant bands of Mr of 32–35 kDa and 40–45 kDa. The saliva immunoblot test exhibits a specificity of 100% and a sensitivity of 98.5%. Thus, saliva could be used as an alternative, non-invasive means for the detection of specific anti-T. gondii IgG in humans.