Mycotoxins are amongst the most prevalent food contaminants leading to serious health implications for humans and animals. Limiting exposure to them within the population remains a pressing food safety challenge. Prevention and timely detection are crucial for minimising mycotoxin contamination in food and feed. Therefore easy-to-use, rapid, eco-friendly and low-cost screening methods are increasingly implemented. Early-stage end-user engagement in the technological development process aids and guides the research towards increased societal impact. To investigate these end-user needs, the limitations of the currently used methods and the expectations towards a novel approach were mapped among stakeholders in a comprehensive survey. Stakeholders along the food supply chain (agricultural, food processing, retail, food safety and control, laboratories) were identified in five European countries and contacted. A total of 63 participants completed the survey, which was followed-up by an interview. The results of the survey revealed that different end-user groups have different priorities. Important limiting factors for agricultural, processing and retail stakeholders of the current methodologies include the complexity of sample preparation, high cost and time-to-results. Complementing the accredited laboratory tests with a pre-screening device would be especially interesting for agricultural producers (87.5% of the participants are interested) and food processors (80%), since there is an increasing demand for on-site detection of contamination. On the other hand, food control authorities and commercial laboratories indicated higher priority towards low quantification limits and multi-compound methods. The time to get the results was found to be more important than the testing cost (important aspect for 74.6 versus 66.7%). Overall, the findings of this study are critical input for end-user-targeted development of novel mycotoxin detection platforms.