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标题：Multiplex Recombinase Polymerase Amplification Assay for the Simultaneous Detection of Three Foodborne Pathogens in Seafood
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作者：Biao Ma ; Jiali Li ; Kai Chen 等
期刊名称：Foods
电子版ISSN：2304-8158
出版年度：2020
卷号：9
期号：3
页码：278-292
DOI：10.3390/foods9030278
出版社：MDPI Publishing
摘要：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.
关键词：recombinase polymerase amplification; lateral flow dipstick; multiplex detection; quantitative detection; Staphylococcus aureus ; Vibrio parahaemolyticus ; Salmonella Enteritidis recombinase polymerase amplification ; lateral flow dipstick ; multiplex detection ; quantitative detection ; Staphylococcus aureus ; Vibrio parahaemolyticus ; Salmonella Enteritidis