An Integrated System Combining Filter-Assisted Sample Preparation and Colorimetric Biosensing for Rapid Pathogen Detection in Complex Food Matrices

Climate change increases microbial contamination risks in food, highlighting the need for real-time biosensors. However, food residues often interfere with detection signals, limiting the direct application. An integrated system of filter-assisted sample preparation (FASP) and an immunoassay-based colorimetric biosensor offers the rapid and simple on-site detection of foodborne pathogens in complex food matrices. The accuracy and stability of biosensor analysis were ensured via filter-assisted preprocessing, which separated food residues from bacteria. The system was applied to various food matrices, including vegetables, meats, and cheese brine, using samples spiked at contamination levels ranging from 10² to 10³ CFU per 25 g, thereby demonstrating broad applicability. Bacterial recovery varied by food matrix, with vegetables showing a 1-log reduction and meats, melon, and cheese brine showing a 2-log reduction relative to the initial inoculum. A detection limit of 10¹ CFU/mL was achieved for <i>Escherichia coli</i> O157:H7, <i>Salmonella</i> Typhimurium, and <i>Listeria monocytogenes</i> in the final preprocessed sample solutions. Sample preparation took under 3 min, and detection was completed within 2 h under stationary conditions. This approach enables rapid pathogen detection in various food matrices without the need for special reading devices, contributing to food safety as a real-time, rapid-response food biosensor.