Improving electrochemical aptasensor sensitivity for Bacillus cereus spore detection in food safety applications

Rapid detection of Bacillus cereus spores is essential for preventing food contamination and spoilage. Many existing methods detect B. cereus vegetative cells rather than spores and cannot be applied directly to foods. Here, we present a combination of aptamers targeting different moieties on the surface of B. cereus spores with rapid electrochemical detection. When DNA aptamers, previously selected for B. cereus spores, were immobilized on screen-printed gold electrodes, they exhibited higher binding capacity than individual aptamers, suggesting a synergistic effect. Additionally, the mixture of rhodamine-labelled aptamers enabled spatial fluorescent visualization of the B. cereus endospore structure, confirming the increased binding efficiency. The electrochemical aptasensor based on three aptamers exhibited a wide dynamic range (10 2 –10 7 CFU/mL) and low limit of detection (∼1 CFU/mL) using just 15 μL of sample. Validation in spiked salad, using direct spore sensing in rinse water and comparison with the culturing method, confirmed its sensitivity and specificity. These combined aptamer approaches, achieving rapid (15 min) and single-step detection may also be suitable for detecting other foodborne pathogens. • A new electrochemical apta-sensor was developed for detection of B. cereus spores. • The triple-aptamer biosensor showed higher signal gain than single ones. • Ready-to-eat salad was used as a matrix for spore quantification. • The assay time was only 15 min.