Specific Detection of Five Bacterial Foodborne Pathogens by Oligonucleotide Macroarray
Keywords:
Multiplex PCR (m-PCR), Oligonucleotide array, Probes, Foodborne pathogensAbstract
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.
References
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Whyte, P., Gill, K.M., Collins, J.D. and Gormley, E. 2002. The prevalence and PCR detection of Salmonella contamination in raw poultry. Veterinary Microbiology. 89: 53–60.
Yeh, K.S., Chen, T.H., Liao, C.W., Chang, C.S. and Lo, H.C. 2002. PCR amplification of the Salmonella Typhimurium fimY gene sequence to detect the Salmonella species. International Journal of Food Microbiology. 78: 227–234.
Yoo, S.M., Keum, K.C., Yoo, S.Y., Choi, J.Y., Chang, K.H., Yoo, N.C., Yoo, W.M., Kim, J.M., Lee, D. and Lee, S.Y. 2004. Development of DNA microarray for pathogen detection. Biotechnology and Bioprocess Engineering. 9: 93–99.
Bartoszewicz, M., Hansen, B.M. and Swiecicka, I. 2008. The members of the Bacillus cereus group are commonly present contaminants of fresh and heat-treated milk. Food Microbiology. 25: 588–596.
Bianchi, D.M., Barbaro, A., Gallina, S., Vitale, N., Chiavacci, L., Caramelli, M. and Decastelli, L. 2013. Monitoring of foodborne pathogenic bacteria in vending machine raw milk in Piedmont, Italy. Food Control. 32: 435–439.
Chen, J. and Griffiths, M.W. 1998. PCR differentiation of Escherichia coli from other gram-negative bacteria using primers derived from the nucleotide sequences flanking the gene encoding the universal stress protein. Letters in Applied Microbiology. 27(6): 369–371.
Chitov, T., Dispan, R. and Kasinrerk, W. 2008. Incidence and diarrhegenic potential of Bacillus cereus in pasteurized milk and cereal products in Thailand. Journal of Food Safety. 28(4): 467–481.
Chou, H.H., Hsia, A.P., Mooney, D.L. and Schnable, P.S. 2004. PICKY: oligo microarray design for large genomes. Bioinformatics. 20(17): 2893–2902.
Chye, F.Y., Abdullah, A. and Ayo, M.K. 2004. Bacteriological quality and safety of raw milk in Malaysia. Food Microbiology. 21: 535–541.
Fusco, V., Quero, G.M., Morea, M., Blaiotta, G. and Visconti, A. 2011. Rapid and reliable identification of Staphylococcus aureus harbouring the enterotoxin gene cluster (egc) and quantitative detection in raw milk by real time PCR. International Journal of Food Microbiology. 144: 528–537.
Gauthier, M. and Blais, B.W. 2003. Comparison of different approaches for the incorporation of non-radioactive labels into polymerase chain reaction products. Biotechnology Letters. 25: 1369–1374.
Germini, A., Masola, A., Carnevali, P. and Marchelli, R. 2009. Simultaneous detection of Escherichia coli O175:H7, Salmonella spp., and Listeria monocytogenes by multiplex PCR. Food Control. 20(8): 733–738.
He, Y., Dai, J. and Li, Y.F. 2011. Simultaneous and rapid detection of enteric pathogens from raw milk by multiplex PCR. Journal of Microbiology and Biotechnology. 27:2597–2602.
Hussain, M., Eiff, C.V., Sinha, B., Joost, I., Herrmann, M., Peters, G. and Becker, K. 2008. eap gene as novel target for specific identification of Staphylococcus aureus. Journal of Clinical Microbiology. 46(2): 470-476.
Jami, S., Jamshidi, A. and Khanzadi, S. 2010. The presence of Listeria monocytogenes in raw milk samples in Mashhad. Iranian Journal of Veterinary Research. 11(4): 363–367.
Kanungpean, D., Rujit, J., Chanda, J., Boonyaprapa, N., Tonchotiwetch, P., Mahanil, W, Tain, A. andIntarapuk, A. 2014. The quality testing of raw goat’s milk in Nongchok District, Bangkok. Journal of Mahanakorn Veterinary Medicine. 9(2): 83–88.
Kim, D.H., Lee, B.K., Kim, Y.D., Rhee, S.K. and Kim, Y.C. 2010. Detection of representative enteropathogenic bacteria, Vibrio spp., pathogenic Escherichia coli, Salmonella spp., Shigella spp., and Yersinia enterocolitica, using a virulence factor gene-based oligonucleotide microarray. Journal of Microbiology. 48(5): 682–688.
Kupradit, C., Innok, S., Woraratphoka, J. and Ketudat-Cairns, M. 2017. Novel multiplex PCR assay for rapid detection of five bacterial foodborne pathogens. Suranaree Journal of Science and Technology. 24(1): 41–50.
Kupradit, C., Rodtong, S. and Ketudat-Cairns, M. 2013a. Development of a DNA macroarray for simultaneous detection of multiple foodborne pathogenic bacteria in fresh chicken meat. World Journal of Microbiology and Biotechnology. 29(12): 2281–2291.
Kupradit, C., Ruamkuson, D., Rodtong, S. and Ketudat-Cairns, M. 2013b. Novel multiplex polymerase chain reaction and an oligonucleotide array for specific detection of the dominant foodborne bacterial pathogens in chicken meat. African Journal of Microbiology Research. 7(24): 3085–3095.
Kupradit, C., Ruamkuson, D., Rodtong, S. and Ketudat-Cairns, M. 2014. Oligonucleotide macroarray for specific detection of bacterial foodborne pathogens. Chiang Mai Journal of Science. 41(5.1): 957–969.
Li, Y., Zhuang, S. and Mustapha, A. 2005. Application of a multiplex PCR for the simultaneous detection of Escherichia coli O157:H7, Salmonella and Shigella in raw and ready-to-eat meat products. Meat Science. 71: 402–406.
Nierop, W.V., Dusé, A.G., Marais, E., Aithma, N., Thothobolo, N., Kassel, M. Stewart, R., Potgieter, A., Fernandes, B., Galpin, J.S. and Bloomfield, S.F. 2005. Contamination of chicken carcasses in Gauteng, South Africa, by Salmonella, Listeria monocytogenes and Campylobacter. International Journal of Food Microbiology. 99: 1–6.
Nugen, S.R. and Baeumner, A.J. 2008. Trends and opportunities in food pathogen detection. Analytical and Bioanalytical Chemistry. 391: 451–454.
Ombarak, R.A., Hinenoya, A., Awasthi, S.P., Iguchi, A., Shima, A., Elbagory, A.R.M. and Yamasaki, S. 2016. Prevalence and pathogenic potential of Escherichia coli isolates from raw milk and raw milk cheese in Egypt. International Journal of Food Microbiology. 221: 69–76.
Padungtod, P. and Kaneene, J.B. 2006. Salmonella in food animals and humans in northern Thailand. International Journal of Food Microbiology. 108: 346–354.
Settanni, L. and Corsetti, A. 2007. The use of multiplex PCR to detect and differentiate food- and beverage-associated microorganisms: A review. Journal of Microbiological Methods. 69: 1–22.
Severgnini, M., Cremonesi, P., Consolandi, C., De Bellis, G. and Castiglioni, B. 2011. Advances in DNA microarray technology for the detection of foodborne pathogens. Food and Bioprocess Technology. 4: 936–953.
United States Food and Drug Administration. 1998. Bacteriological analytical manual / Food and Drug Administration, 8th ed, AOAC International, Gaithersburg, MD.
Van Kessel, J.S., Karns, J.S., Gorski, L., McCluskey, B.J. and Perdue, M.L. 2004. Prevalence of Salmonellae, Listeria monocytogenes and fecal coliforms in bulk tank milk on US Dairies. Journal of Dairy Science. 87: 2822–2830.
Whyte, P., Gill, K.M., Collins, J.D. and Gormley, E. 2002. The prevalence and PCR detection of Salmonella contamination in raw poultry. Veterinary Microbiology. 89: 53–60.
Yeh, K.S., Chen, T.H., Liao, C.W., Chang, C.S. and Lo, H.C. 2002. PCR amplification of the Salmonella Typhimurium fimY gene sequence to detect the Salmonella species. International Journal of Food Microbiology. 78: 227–234.
Yoo, S.M., Keum, K.C., Yoo, S.Y., Choi, J.Y., Chang, K.H., Yoo, N.C., Yoo, W.M., Kim, J.M., Lee, D. and Lee, S.Y. 2004. Development of DNA microarray for pathogen detection. Biotechnology and Bioprocess Engineering. 9: 93–99.
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Published
2018-08-21
How to Cite
Kupradit, Chanida, and Mariena Ketudat-Cairns. 2018. “Specific Detection of Five Bacterial Foodborne Pathogens by Oligonucleotide Macroarray”. Food and Applied Bioscience Journal 7 (1):1-17. https://li01.tci-thaijo.org/index.php/fabjournal/article/view/127943.
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Food Processing and Engineering
