Rapid, sensitive, and selective detection of live pathogens remains a key priority for quality control and risk assessment. While conventional methods often require complicated workflows, costly reagents, lab equipment, and are time-consuming, rendering them inadequate for field testing and lowresource settings. Increased attention has been drawn to developing alternative low-cost and rapid methods to detect on-site live pathogens in different environmental matrices. Among them, microfluidic devices that integrate various laboratory functions in a miniaturized manner have proven to be a promising tool for the rapid and sensitive detection of pathogens. Herein, the development of microfluidic devices specifically designed for the detection of live pathogens is discussed along a concise summary of novel microfluidics systems recently developed, contrasted to conventional methods regarding assay time, the limit of detection, and target organisms. These include a variety of micro total analysis systems (µTAS) and micro fluidic paper-based analytical devices (µPADs) in combination with molecular methods and traditional live cell detection techniques, such as cell culture, DNA intercalating dyes, resazurin, and immobilized bioreceptors (e.g., aptamers and capture antibodies). Furthermore, insights on the future perspectives of microfluidics for live pathogen detection with a highlight on the rapid and low-cost method development for field testing are provided.

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