Real-Time PCR Assays for Quantification of Trypanosoma cruzi in Food Samples: A Tool for Surveillance of Oral Chagas Disease Outbreaks.

Chagas disease (ChD), caused by Trypanosoma cruzi, remains a significant public health concern in Latin America, despite advances in control measures. While traditionally associated with vector-borne transmission, oral transmission has emerged as the predominant route for acute infections, particularly through contaminated food and beverages. Outbreaks have been reported across Latin America, with Brazil accounting for a significant number of cases, especially in the Amazon region. Consumption of açai pulp and sugarcane juice has been frequently implicated in outbreaks of acute Chagas disease orally transmitted in Brazil. The açai palm-tree fruit is one of the most widely marketed Amazonian fruits, both in Brazil and internationally. Thus, it is essential to incorporate procedures into the production process of açaí juice and pulp that ensure effective hygiene and quality control. Hence, the molecular diagnostic based on qPCR and RT-qPCR assays can be powerful tools for surveillance and control of orally-transmitted ChD. In this chapter, we describe a real-time PCR methodology to the detection and quantification of T. cruzi DNA and RNA, capable of differentiating viable parasites in the food samples.