Specific Detection of Five Bacterial Foodborne Pathogens by Oligonucleotide Macroarray
Keywords:Multiplex PCR (m-PCR), Oligonucleotide array, Probes, Foodborne pathogens
Milk and dairy products can be contaminated with a variety of microorganisms. Thus, a rapid method for simultaneous detection of multiple foodborne pathogens should be considered. In this investigation, a combination of multiplex PCR (m-PCR) and oligonucleotide array hybridization was performed to specifically detect multiple foodborne pathogens. Specific genes, including Enterotoxin FM, uspA, prfA, fimY, and eap genes, were selected as targets for detection of Bacillus cereus, Escherichia coli, Listeria monocytogenes, Salmonella spp., and Staphylococcus aureus, respectively. The suitable probes for specific detection of five foodborne pathogens were selected based on the specificity of detection. The hybridization signals of digoxigenin (DIG) incorporated into the PCR target regions were observed by naked eyes. Differences of hybridization patterns were observed among isolated and reference strains of B. cereus and S. aureus. High accuracies of specific probes, including, probes BC1, BC3 and BC5 (30–100% accuracy); probes EC1–EC4 (83–100% accuracy); probes LM1, LM2, LM4 (87–100% accuracy); probes SA1–SA5 (75–100% accuracy); and probes SM1, SM3 (66–100% accuracy), were selected for detection of B. cereus, E. coli, L. monocytogenes, S. aureus, and Salmonella spp., respectively. For future work, the DNA target of each bacterium will be amplified by PCR followed by hybridization with the suitable probes obtained from this work.
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