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A Gold Nanoparticles Assisted Multiplex PCR Assay for Simultaneous Detection of Salmonella typhimurium, Listeria monocytogenes and Escherichia coli O157:H7

Abstract

Foodborne pathogens are a major cause of foodborne illness, leading to a growing food safety problem in public health. This work aims to develop a novel gold nanoparticles (AuNPs) assisted multiplex PCR assay for rapid, simple and simultaneous detection of Salmonella typhimurium (S. typhimurium), Listeria monocytogenes (L. monocytogenes) and Escherichia coli O157:H7 (E. coli O157:H7), which were the top three foodborne pathogenic bacteria. Flower-shaped AuNPs (F-AuNPs) were used as colorimetric sensor in this assay, based on PCR product can help improve stability of the F-AuNPs in a certain concentration of salt solution. Detection of PCR product can be directly achieved by mixing it with F-AuNPs and NaCl, and the result is read by naked eyes. Results showed that the optimal annealing temperature was 53.1oC to amplify the target three pathogenic strains in multiplex PCR assay, and the optimal concentrations of the primer pairs was 0.4 µM each for L.monocytogenes and E. coli O157:H7, and 0.2 µM each for S. typhimurium. Colorimetric detection limit of PCR products by F-AuNPs was 3.125 ng/µL, and the detection time was approximately 10 min. Simultaneous detection limit of the multiplex PCR method was 10 pg/µL for L. monocytogenes and S. typhimurium, and 50 pg/µL for E. coli O157:H7. Compared with conventional multiplex PCR assay, the F-AuNPs assisted assay is a convenient, rapid and simple visual detection method. The excellent performance of the colorimetric sensor shows the potential application in on-site detection of foodborne pathogenic strains in food samples

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Publication details

The article was received on 22 Oct 2019, accepted on 03 Dec 2019 and first published on 04 Dec 2019


Article type: Paper
DOI: 10.1039/C9AY02282A
Anal. Methods, 2019, Accepted Manuscript

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    A Gold Nanoparticles Assisted Multiplex PCR Assay for Simultaneous Detection of Salmonella typhimurium, Listeria monocytogenes and Escherichia coli O157:H7

    J. Du, S. Wu, L. Niu, J. Li, D. Zhao and Y. Bai, Anal. Methods, 2019, Accepted Manuscript , DOI: 10.1039/C9AY02282A

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