Preventing Chagas disease: a new RT-qPCR method for rapid and specific quantification of viable Trypanosoma cruzi for food safety

Abstract Without standardized methods for rapidly detecting in food matrices viable T. cruzi, foodborne outbreaks remain neglected. In this work, a reverse-transcriptase real-time PCR (RT-qPCR) mRNA-based technique was developed for the rapid and specific detection and quantification of viable Trypanosoma cruzi in acai fruits and juice. The method uses specific primer targeting region on the cyt b gene. The maximum recovery rate of T. cruzi from inoculated acai juice was 82.50 %. The limit of detection and quantification in acai juice was 10 parasites/mL for RT-qPCR (mRNA-based) and qPCR (DNA-based). The RT-qPCR efficiency was estimated at 97.27% with an R2 of 0.994. The RT-qPCR was shown to be able to discriminate between viable and nonviable cells. This method provides a useful tool for rapid assessment of low concentrations of viable T. cruzi in naturally contaminated food samples, and can be applied industrially as a quality and security method.