Simultaneous Detection of Four Main Foodborne Pathogens in Ready-to-Eat Food by Using a Simple and Rapid Multiplex PCR (mPCR) Assay

Aya Boukharouba, Ana González, Miguel García-Ferrús, María Antonia Ferrús and Salut Botella
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Aya Boukharouba: Department of Biotechnology, Centro Avanzado de Microbiología de Alimentos, Universitat Politècnica de València, Camino de Vera, s/n, 46022 Valencia, Spain
Ana González: Department of Biotechnology, Centro Avanzado de Microbiología de Alimentos, Universitat Politècnica de València, Camino de Vera, s/n, 46022 Valencia, Spain
Miguel García-Ferrús: Department of Biotechnology, Centro Avanzado de Microbiología de Alimentos, Universitat Politècnica de València, Camino de Vera, s/n, 46022 Valencia, Spain
María Antonia Ferrús: Department of Biotechnology, Centro Avanzado de Microbiología de Alimentos, Universitat Politècnica de València, Camino de Vera, s/n, 46022 Valencia, Spain
Salut Botella: Department of Biotechnology, Centro Avanzado de Microbiología de Alimentos, Universitat Politècnica de València, Camino de Vera, s/n, 46022 Valencia, Spain

IJERPH, 2022, vol. 19, issue 3, 1-18

Abstract: The increasing consumption of organic or ready-to-eat food may cause serious foodborne disease outbreaks. Developing microbiological culture for detection of food-borne pathogens is time-consuming, expensive, and laborious. Thus, alternative methods such as polymerase chain reaction (PCR) are usually employed for outbreaks investigation. In this work, we aimed to develop a rapid and simple protocol for the simultaneous detection of Escherichia coli ( E coli ), Listeria monocytogenes ( L. monocytogenes ), Staphylococcus aureus ( S. aureus ) and Salmonella enterica ( S. enterica ), by the combination of an enrichment step in a single culture broth and a multiplex PCR (mPCR) assay. The effectiveness of several enrichment media was assessed by culture and PCR. Buffered peptone water (BPW) was selected as the optimum one. Then, mPCR conditions were optimized and applied both to pure co-cultures and artificially inoculated food samples (organic lettuce and minced meat). In the culture medium inoculated at 10 0 CFU/mL, mPCR was able to detect the four microorganisms. When performed on artificially food samples, the mPCR assy was able to detect E. coli , S. enterica , and L. monocytogenes . In conclusion, BPW broth can effectively support the simultaneous growth of E. coli , S. aureus , L. monocytogenes , and S. enterica and could be, thus, used prior to a mPCR detection assay in ready-to-eat food, thereby considerably reducing the time, efforts and costs of analyzes.

Keywords: food safety; detection; multiplex PCR (mPCR); co-enrichment; organic food; ready-to-eat food; Escherichia coli; Listeria monocytogenes; Staphylococcus aureus; Salmonella enterica (search for similar items in EconPapers)
JEL-codes: I I1 I3 Q Q5 (search for similar items in EconPapers)
Date: 2022
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