Development and Evaluation of a Multiplex PCR for Simultaneous Detection of Five Foodborne Pathogens

Authors: Thuy Trang Nguyen, Vo Van Giau, Tuong Kha Vo

Foodborne pathogens present serious concerns to human health and can even lead to fatalities. The gold standard for pathogen identification – bacterial culture – is costly and time consuming. A cheaper and quicker alternative will benefit in controlling food safety. In this study, we developed a multiplex-PCR protocol for simultaneous detection of five Foodborne pathogens including Escherichia coli O157:H7, Staphylococcus aureus, Salmonella spp., Listeria monocytogenes, and Vibrio cholerae, based on five genes stx1, femA, invA, iap, và ctxA, respectively. Specific primers for multiplex PCR amplification of the stx (Shiga-like toxin), nuc (thermo nuclease), inv A (invasion protein A), iap (invasive associative protein), and ctx A (cholera toxin A) genes that were established to amplify simultaneous detection of the target pathogens. The assay was also validated for its specificity, sensitivity, and applied to test some spiked food samples. The results showed the products expected multiplex PCR fragments of approximately 112, 244, 301, 453, 518 and 720bp for S. aureus, Salmonella spp. V. cholera, L. monocytogenes, E. coli O157:H7 and 16S rRNA, respectively. The assay was specific to the targeted pathogens and was sufficiently sensitive and robust to effectively analyze market samples. The whole process took less than 24 h to complete indicating that the assay is suitable for reliable and rapid identification of these five foodborne pathogens, which could be suitable in microbial epidemiology investigation. You can submit your Manuscripts at: 

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[v1] 2017-08-28 04:17:55

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