Chagas disease has the highest prevalence of any parasitic disease in the Americas, affecting 6–7 million people. Conventional diagnosis requires a well-equipped laboratory with experienced personnel. The development of new diagnostic tools that are easy to use and adapted to the reality of affected populations and health systems is still a significant challenge. The main objective of this study was to measure Trypanosoma cruzi infection status using saliva samples of infected subjects. Blood and saliva samples from 20 T. cruzi-seropositive individuals and 10 controls were tested for T. cruzi infection using two different commercial serological tests. We have shown that detection of T. cruzi infection is possible using saliva samples, supporting the potential use of saliva to diagnose Chagas disease in humans. This method could provide a simple, low-cost but effective tool for the diagnosis of T. cruzi infection. Its noninvasive nature makes it particularly well suited for endemic areas.