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http://dx.doi.org/10.5851/kosfa.2012.32.2.241

Simultaneous Detection of 10 Foodborne Pathogens using Capillary Electrophoresis-Based Single Strand Conformation Polymorphism  

Oh, Mi-Hwa (National Institute of Animal Science, Rural Development Administration)
Hwang, Hee-Sung (Department of Chemical Engineering, Pohang University of Science and Technology)
Chung, Bo-Ram (Department of Chemical Engineering, Pohang University of Science and Technology)
Paik, Hyun-Dong (Department of Food Science and Biotechnology of Animal Resource, Konkuk University)
Han, Sang-Ha (National Institute of Animal Science, Rural Development Administration)
Kang, Sun-Moon (National Institute of Animal Science, Rural Development Administration)
Ham, Jun-Sang (National Institute of Animal Science, Rural Development Administration)
Kim, Hyoun-Wook (National Institute of Animal Science, Rural Development Administration)
Seol, Kuk-Hwan (National Institute of Animal Science, Rural Development Administration)
Jang, Ae-Ra (Department of Animal Products and Food Science, Kangwon National University)
Jung, Gyoo-Yeol (Department of Chemical Engineering, Pohang University of Science and Technology)
Publication Information
Food Science of Animal Resources / v.32, no.2, 2012 , pp. 241-246 More about this Journal
Abstract
This report outlines the development of a rapid, simple, and sensitive detection system for pathogenic bacteria using a capillary electrophoresis-based, single strand conformation polymorphism (CE-SSCP) combined with PCR. We demonstrate that this method, used with primers targeting the V4 region of the16S rRNA gene, is capable of the simultaneous detection of 10 microbes that could be associated with foodborne illness, caused by animal-derived foods: Salmonella enterica, Listeria monocytogenes, Escherichia coli O157:H7, Campylobacter jejuni, Staphylococcus aureus, Bacillus cereus, Clostridium perfringens, Yersinia enterocolitica, Vibrio parahaemolyticus, and Enterobacter sakazakii. The traditional detection techniques are time-consuming and labor-intensive, due to the necessary task of separate cultivation of each target species. As such, the CE-SSCP-PCR method, that we have developed, has the potential to diagnose pathogens rapidly, unlike the traditional technique, in order to prevent foodborne illness in a much more efficient manner.
Keywords
CE; SSCP; PCR; simultaneous detection; foodborne pathogens;
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1 Wesche, H., Xiao, S. H., and Young, S. W. (2005) High throughput screening for protein kinase inhibitors. Comb. Chem. High Through. Screen 8, 181-195.   DOI   ScienceOn
2 Zheng, W., Spencer, R. H., and Kiss, L. (2004) High throughput assay technologies for ion channel drug discovery. Assay. Drug Dev. Ttechnol. 2, 543-552.   DOI   ScienceOn
3 Zinger, L., Gury, J., Giraud, F., Krivobok, S., Gielly, L., Taberlet, P., and Geremia, R. A. (2007) Improvements of polymerase chain reaction and capillary electrophoresis singlestrand conformation polymorphism methods in microbial ecology toward a high-throughput method for microbial diversity studies in soil. Microb. Ecol. 54, 203-216.   DOI
4 Jos Van, A., Francis, D. S., Gaetan, M., Adel, B., Anne, N., and Sabine, L. (2001) Outbreak of necrotizing Enterocolitis associated with Enterobacter sakazakii in powdered milk formula. J. Clin. Microbiol. 39, 293-297.   DOI   ScienceOn
5 Oh, M. H. and Cox, J. M. (2009) Toxigenic bacilli associated with food poisoning. Food Sci. Biotechnol. 18, 594-603.
6 Kullen, M. J., Sanozky-Dawes, R. B., Crowell, D. C., and Klaenhammer, T. R. (2000) Use of the DNA sequence of variable regions of the 16S rRNA gene for rapid and accurate identification of bacteria in the Lactobacillus acidophilus complex. J. Appl. Microbiol. 89, 511-516.   DOI   ScienceOn
7 Larsen, L. A., Jespersgaard, C., and Andersen, P. S. (2007) Single- strand conformation polymorphism analysis using capillary array electrophoresis for large-scale mutation detection. Nat. Protoc. 2, 1458-1466.   DOI   ScienceOn
8 McNally, A., Cheasty, T., Fearnley, C., Dalziel, R. W., Paiba, G. A., Manning, G., and Newell, D. G. (2004). Comparison of the biotypes of Yersinia enterocolitica isolated from pigs, cattle and sheep at slaughter and from humans with yersiniosis in Great Britain during 1999-2000. Lett. Appl. Microbiol. 39, 103-108.   DOI   ScienceOn
9 Oh, M. H., Park, Y. S., Paek, S. H., Shin, G. W., Kim, H. Y., Jung, G. Y., and Oh, S. (2009) Simultaneous identification of seven food-borne pathogens and Escherichia coli (pathogenic and non-pathogenic) using capillary electrophoresisbased single-strand conformation polymorphism coupled with multiplex PCR. J. Food Prot. 72, 1262-1266.   DOI
10 Sakamoto, M., Takeuchi, Y., Umeda, M., Ishikawa, I and Benno, Y. (2001) Rapid detection and quantification of five periodontopathic bacteria by real-time PCR. Microbiol. Immunol. 45, 39-44   DOI
11 Shin, G. W., Cho, Y. S., Hwang, H. S., Park, J. H., and Jung, G. Y. (2008) A two-step quantitative pathogen detection system based on capillary electrophoresis. Anal. Biochem. 383, 31-37.   DOI   ScienceOn
12 Shin, G. W., Hwang, H. S., Seo, S. W., Oh, M. H., Ryu, C. Y., Salvo, C. J., Feldman, S., Doh, J., and Jung, G. Y. (2010) A novel pathogen detection system based copolymer matrix. J. Sep. Sci. 33, 1639-1643.   DOI   ScienceOn
13 Claesson, M. J., Wang, Q., O'Sullivan, O., Greene-Diniz, R., Cole, J. R., Ross, R. P., and O'Toole, P. W. (2010) Comparison of two next-generation sequencing technologies for resolving highly complex microbiota composition using tandem variable 16S rRNA gene regions. Nucl. Acids Res. 38, 1-13.   DOI
14 Altekruse, S. F., Stern, N. J., Fields, P. I., and Swerdlow, D. L. (1999) Campylobacter jejuni - An emerging foodborne pathogen. Emer. Infec. Dis. 5, 28-35.   DOI   ScienceOn
15 Andersen, P. S., Jespersgaard, C., Vuust, J., Christiansen, M., and Larsen, L. A. (2003) Capillary electrophoresis-based single strand DNA conformation analysis in high-throughput mutation screening. Hum. Mutat. 21, 455-465.   DOI   ScienceOn
16 Animal Plant and Fisheries Quarantine and Inspection Agency. Standard for processing and ingredients of specifications livestock products. Available from: http://www.nvrqs.go.kr/Ex_Work/Scflp/Livestock/View.asp. Accessed Jun. 21, 2011.
17 Gillman, L. M., Gunton, J., Turenne, C. Y., Wolfe, J., and Kabani, A. M. (2001) Identification of Mycobacterium species by multiple-fluorescence PCR-single-strand conformation polymorphism analysis of the 16S rRNA gene. J. Clin. Microbiol. 39, 3085-3091.   DOI
18 Gracias, K. S. and Mackillip, J. L. (2004) A review of conventional detection and enumeration methods for pathogenic bacteria in food. Can. J. Microbiol. 50, 883-890.   DOI   ScienceOn
19 Iversen, C. and Forsythe, S. (2003) Risk profile of Enterobacter sakazakii, an emergent pathogen associated with infant milk formula. Trend. Food Sci. Technol. 14, 443-454.   DOI   ScienceOn