Abstract
Foodborne pathogens can cause foodborne illness. In reality, one food sample may carry more than one pathogen. A rapid, sensitive, and multiple target method for bacteria detection is crucial in food safety. For the simultaneous detection of Staphylococcus aureus, Vibrio parahaemolyticus, and Salmonella Enteritidis, multi-objective recombinase polymerase amplification (RPA) combined with a lateral flow dipstick (LFD) was developed in this study. The whole process, including amplification and reading, can be completed in 15 min at 37 °C. The detection limits were 2.6 × 101 CFU/mL for Staphylococcus aureus, 7.6 × 101 CFU/mL for Vibrio parahaemolyticus, and 1.29 × 101 CFU/mL for Salmonella Enteritidis. Moreover, colored signal intensities on test lines were measured by a test strip reader to achieve quantitative detection for Staphylococcus aureus (R2 = 0.9903), Vibrio parahaemolyticus (R2 = 0.9928), and Salmonella Enteritidis (R2 = 0.9945). In addition, the method demonstrated good recoveries (92.00%–107.95%) in the testing of spiked food samples. Therefore, the multiplex LFD-RPA assay is a feasible method for the rapid, sensitive, and quantitative detection of bacterial pathogens in seafood.
Highlights
Seafood products have become increasingly popular among consumers across the world
We report a combination of isothermal recombinase polymerase amplification (RPA) and lateral flow dipstick (LFD) for the simultaneous detection of Staphylococcus aureus, Vibrio parahaemolyticus, and Salmonella Enteritidis
Primers for the Vibrio parahaemolyticus fragment were decreased to 150 nM, and a range of concentrations of the primers for the Staphylococcus aureus and Salmonella Enteritidis fragments
Summary
Seafood products have become increasingly popular among consumers across the world. Seafood has frequently been associated with foodborne illness because it is contaminated with pathogens during cultivation, handling, and processing. Foodborne pathogens seriously threaten human health and can cause various diseases and even death. The World Health Organization released a report in 2015 stating that 70% of foodborne diseases were caused by pathogenic microorganisms. The five most prevalent foodborne pathogens are Staphylococcus aureus, Vibrio parahaemolyticus, Salmonella, Listeria monocytogenes, and Escherichia coli O157:H7 [1]. Staphylococcus aureus, Vibrio parahaemolyticus, and Salmonella are important foodborne pathogens in seafood products [2,3]
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