Fast molecular methods for the detection of spoilage fungi (in food products)

Resumo

In the present thesis, the detection of spoilage and mycotoxigenic fungi in different food matrices was achieved in 24-48 h, providing a faster alternative to the conventional culture-based techniques which require up to 7 days from sampling to result. Consequently, the developed methods present an interesting alternative for the food industry resulting in reduction of costs associated with lengthy analyses and product recalls. Small modifications of the proposed methodology including enrichment time and temperature as well as sample size have proven to contribute to higher sensitivity and/or specificity which gives the developed methodology the required flexibility to fit the needs of the food industry. Furthermore, the different methods showcased different advantages. In particular, the real-time PCR assays have proven to be more sensitive; however, they require a real-time thermocycler in order to be performed. On the other hand, the isothermal amplification techniques coupled with naked-eye detection could be used for POC testing and early screening since they only require a heating device; providing in many cases a comparable sensitivity to the qPCR assays.