Mesoporous Silica\nNanoparticles-Enhanced Microarray\nTechnology for Highly Sensitive Simultaneous Detection of Multiplex\nFoodborne Pathogens

Abstract

Ensuring food safety is paramount for the food industry\nand global\nhealth concerns. In this study, we have developed a method for the\ndetection of prevalent foodborne pathogenic bacteria, including Escherichia coli, <i>Salmonella</i> spp., <i>Listeria</i> spp., <i>Shigella</i> spp., <i>Campylobacter</i> spp., <i>Clostridium</i> spp., and <i>Vibrio</i> spp., utilizing antibody-aptamer arrays. To enhance the fluorescence\nsignals on the microarray, the mesoporous silica nanoparticles (MSNs)\nconjugated with fluorescein, streptavidin, and seven detection antibodies-biotin\nwere employed, forming fluorescein doped mesoporous silica nanoparticles\nconjugated with detection antibodies (MSNs-Flu-SA-Abs) complexes.\nThe array pattern was designed for easy readability and enabled the\nsimultaneous detection of all seven foodborne pathogens, referred\nto as the 7FP-biochip. Following the optimization of MSNs-Flu-SA-Abs\ncomplexes attachment and enhancement of the detection signal in fluorescent\nimmunoassays, a high level of sensitivity was achieved. The detection\nlimits for the seven pathogens in both buffer and food samples were\n10² CFU/mL through visual screening, with fluorescent intensity\nquantification achieving levels as low as 20–34 CFU/g were\nachieved on the antibody-aptamer arrays. Our antibody-aptamer array\noffers several advantages, including significantly reduced nonspecific\nbinding with no cross-reaction between bacteria. Importantly, our\nplatform detection exhibited no cross-reactivity among the tested\nbacteria in this study. The multiplex detection of foodborne pathogens\nin canned tuna samples with spiked bacteria was successfully demonstrated\nin real food measurements. In conclusion, our study presents a promising\nmethod for detecting multiple foodborne pathogens simultaneously.\nWith its high sensitivity and specificity, the developed antibody-aptamer\narray holds great potential for enhancing food safety and public health.