Toxoplasmosis is an anthropozoonotic disease endemic world-wide, caused by the apicomplexan Toxoplasma gondii. Although the course of infection is generally benign, it can cause significant morbidity and mortality in the developing fetus and in immunocompromised individuals. Biological diagnosis classically relies upon serology and direct detection of the parasite by inoculation to laboratory animals. In the past decade, the use of the polymerase chain reaction (PCR) has made a significant improvement in both the prenatal diagnosis of congenital toxoplasmosis and the detection of acute disease in the immunocompromised patient. Nevertheless, like many 'in-house' PCR assays, the PCR-Toxoplasma suffers from lack of standardization and variable performance according to the laboratory. A wide variety of primers has been used in different assays, but few comparative tests have been performed. Moreover, in contrast to other parasitic diseases, PCR-Toxoplasma has not yet attained a sufficient level of sensitivity, regardless of the clinical condition considered. These drawbacks are discussed, together with the undoubted gain that PCR has brought to this difficult diagnosis.