•Trypanosoma cruzi infection is endemic in the Bolivian Chaco.•Childhood T. cruzi seroprevalence was found to be 0.22, rising steeply with age.•Persistent transmission persists, as shown by a force of infection (FOI) annual incidence estimate of 0.021.•School-based screening with rapid tests is a practical approach in remote areas.•Current intermittent vector control practices in the Bolivian Chaco are suboptimal. Screening for Trypanosoma cruzi infection was performed amongst children in a rural community in the Bolivian Chaco, an area known for high prevalence. The force of infection (FOI) was estimated. A total of 423 children attending the local school were screened using the Chagas Detect Plus (CDP) rapid test (InBios International, Inc.). CDP-positive specimens were further tested by indirect haemagglutination assay (IHA) and Wiener Recombinante v3.0 ELISA. A catalytic model was used to estimate FOI. Confirmed seroprevalence was 0.22, rising steeply with age. The mean age of seropositive individuals was 13 years. The calculated specificity of the rapid test was 91.9%. The annual incidence estimated from the FOI was 0.021. This study demonstrates persistent transmission and continuing high levels of T. cruzi infection in the Bolivian Chaco, and highlights the practicality of school-based screening.