In xenodiagnosis, the vector of a pathogenic agent is infected to make identification of the parasite easier after a period of multiplications. This enables a diagnosis if the strain of the pathogen possesses a low virulence or if very low numbers of parasites occur in the blood, for example Trypanosoma cruzi in the chronic phase of Chagas disease. In 1914, 5 years after the discovery of T. cruzi in the triatomines, its blood-sucking vectors, these were used for the first time as a diagnostic tool of this disease. For many decades thereafter xenodiagnosis represented the golden standard. Meanwhile serological and molecular biological methods are preferably used for diagnosis today, but xenodiagnosis and in vitro cultivation are still the best methods for a proof of the living parasites or for the isolation of the flagellate. In this context, the different methodological variations and the disadvantages and advantages of xenodiagnosis are discussed in our review.