Abstract：Pathogenic bacteria pose a serious threat to human health, and an excessively long detection time may cause patients to miss the optimal treatment opportunity. Therefore, rapid and accurate bacterial detection methods are highly significant for the diagnosis of diseases. This study aimed to develop a simple and portable detection platform for the highly sensitive colorimetric detection of P. aeruginosa, S. aureus, and E. coli. The functional NFM with a double-layer structure was used as the research material. One layer consisted of a nanofiber film loaded with Fe(III), while the other layer was a hydrophilic polymer.The functional NFM demonstrated a detection limit of 10 CFU/mL for pathogenic bacteria, with a detection time of only 10 min. This platform showed great potential in environmental and food safety detection.