title: Simultaneous Identification of 13 Foodborne Pathogens by Using Capillary Electrophoresis-Single Strand Conformation Polymorphism Coupled with Multiplex Ligation-Dependent Probe Amplification and Its Application in Foods.
abstract: Capillary electrophoresis-single strand conformation polymorphism (CE-SSCP) coupled with stuffer-free multiplex ligation-dependent probe amplification (MLPA) was developed to identify 13 species of foodborne pathogens simultaneously. Species-specific MLPA probes were designed for nine of these species. These probes were targeted to the groEL, glyA, MMS, tuf, inv, ipaH, nuc, vvh, and 16S rRNA genes, which corresponded to Bacillus cereus, Campylobacter coli, Cronobacter sakazakii, Enterococcus spp., Salmonella spp., Shigella spp., Staphylococcus aureus, Vibrio vulnificus, and Yersinia enterocolitica, respectively. MLPA probes that had been previously developed by our laboratory were used for the other four species (Campylobacter jejuni, Clostridium perfringens, Escherichia coli O157:H7, and Listeria monocytogenes). The CE-SSCP method was optimized to identify all 13 foodborne microbes simultaneously in a single electrogram, in which 50-500 pg genomic DNA was detected per microbe. Twelve species were detected from animal-derived food samples (specifically, milk and sliced ham) that had been artificially inoculated with 12 of the foodborne pathogens, excluding V. vulnificus, which is not usually associated with animal foods. The method developed here could be used as an early warning system for outbreaks of foodborne diseases associated with animal-derived foods in the food industry.
authors: 1 National Institute of Animal Science , Rural Development Administration, Wanju-gun, Korea.
authors: So-Young
authors: SY
authors: Kim
authors: 2 Department of Chemical Engineering, Pohang University of Science and Technology , Pohang, Korea.
authors: Boram
authors: B
authors: Chung
authors: 1 National Institute of Animal Science , Rural Development Administration, Wanju-gun, Korea.
authors: Jin-Hee
authors: JH
authors: Chang
authors: 2 Department of Chemical Engineering, Pohang University of Science and Technology , Pohang, Korea.|3 School of Interdisciplinary Bioscience and Bioengineering, Pohang University of Science and Technology , Pohang, Korea.
authors: Gyoo Yeol
authors: GY
authors: Jung
authors: 1 National Institute of Animal Science , Rural Development Administration, Wanju-gun, Korea.
authors: Hyoun Wook
authors: HW
authors: Kim
authors: 1 National Institute of Animal Science , Rural Development Administration, Wanju-gun, Korea.
authors: Beom-Young
authors: BY
authors: Park
authors: 4 Department of Food Science and Technology, College of Engineering, Ewha Womans University , Seoul, Korea.
authors: Sang Suk
authors: SS
authors: Oh
authors: 1 National Institute of Animal Science , Rural Development Administration, Wanju-gun, Korea.
authors: Mi-Hwa
authors: MH
authors: Oh
mesh_terms: D000818:Animals; D002417:Cattle; D015342:DNA Probes / Q000032:analysis / Q000378:metabolism; D004269:DNA, Bacterial / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D019075:Electrophoresis, Capillary; D005513:Food Inspection / Q000379:methods*; D005517:Foodborne Diseases / Q000382:microbiology*; D005819:Genetic Markers; D006090:Gram-Negative Bacteria / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D006094:Gram-Positive Bacteria / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D006801:Humans; D057230:Limit of Detection; D008460:Meat / Q000382:microbiology; D008892:Milk / Q000382:microbiology; D058889:Molecular Typing / Q000379:methods*; D060885:Multiplex Polymerase Chain Reaction; D018807:Polymorphism, Single-Stranded Conformational; D015203:Reproducibility of Results; D056910:Republic of Korea; D034421:Sus scrofa
chemical_list: D015342:DNA Probes; D004269:DNA, Bacterial; D005819:Genetic Markers
keywords: Capillary electrophoresis; foodborne pathogens; simultaneous detection; simultaneous identification