Fast, Affordable and multiplexed foodborne pathogen detection on miniaturized devices

Resumo

Food poisoning is a global public health concern affecting, not only developing, but also developed countries, where several pathogens are responsible for these foodborne diseases causing high number of hospitalization and death. The objective of this thesis was to develop an improved methodology for the multiplex detection of Salmonella spp., Shiga Toxin-producing E. coli (STEC) and L. monocytogenes, based in DNA detection, and its integration in a miniaturized device. To achieve this goal, the different steps of the analysis, including the sample pre-treatment, DNA amplification and visualization of the results were addressed in order to reduce the time of analysis, reduce the cost, and allow naked-eye detection. The methodology developed in this project allowed for the multiplex detection of the three targets, reducing the time of analysis to only 9 h vs 7 days by traditional approaches with similar sensitivity and specificity to qPCR. Furthermore, the possibility to integrate the amplification step in a miniaturize device opens the door for the development of Point-of-Care systems.