Multiplex Recombinase Polymerase Amplification Assay for the Simultaneous Detection of Three Foodborne Pathogens in Seafood

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 degrees C. The detection limits were 2.6 x 10(1) CFU/mL for Staphylococcus aureus, 7.6 x 10(1) CFU/mL for Vibrio parahaemolyticus, and 1.29 x 10(1) 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 (R-2 = 0.9903), Vibrio parahaemolyticus (R-2 = 0.9928), and Salmonella Enteritidis (R-2 = 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.

Keywords

recombinase polymerase amplification; lateral flow dipstick; multiplex detection; quantitative detection; Staphylococcus aureus; Vibrio parahaemolyticus; Salmonella Enteritidis

Published in

Foods
2020, Volume: 9, number: 3, article number: 278
Publisher: MDPI

SLU Authors

• Sun, Chuanxin

  • Department of Plant Biology, Swedish University of Agricultural Sciences
    

Sustainable Development Goals

SDG2 Zero hunger


UKÄ Subject classification

Food Science


Publication identifier

DOI: https://doi.org/10.3390/foods9030278

Permanent link to this page (URI)

https://res.slu.se/id/publ/105387