Because of its life cycle, the recovery of Toxoplasma gondii from biological samples is often impracticable. Consequently, a serological diagnosis represents the first and the most widely used approach to defining the stage of infection. The detection of IgG, IgM, IgA, IgE and IgG avidity by different methods offers this opportunity. However, the results may be affected by difficulties in interpretation, as the same antibody pattern may have a different valency, contingent upon subjects and clinical settings, e.g., pregnant women vs. neonates, and treated vs. untreated patients. This review describes the various factors that should be taken into account when performing serological tests for T. gondii, as well as the pitfalls that may be encountered during the interpretative process.