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REP-PCR Genotyping of Four Major Gram-negative Foodborne Bacterial Pathogens  

Jung, Hye-Jin (Department of Food Science and Technology, Chung-Ang University)
Seo, Hyeon-A (Department of Food Science and Technology, Chung-Ang University)
Kim, Young-Joon (Department of Food Science and Technology, Chung-Ang University)
Cho, Joon-Il (Department of Food Science and Technology, Chung-Ang University)
Kim, Keun-Sung (Department of Food Science and Technology, Chung-Ang University)
Publication Information
Korean Journal of Food Science and Technology / v.37, no.4, 2005 , pp. 611-617 More about this Journal
Abstract
Dispersed repetitive DNA elements in genomes of microorganisms differ among and within species. Because distances between repetitive sequences vary depending on bacterial strains, genomic fingerprinting with interspersed repetitive sequence-based probes can be used to distinguish unrelated organisms. Among well-known bacterial repetitive sequences, Repetitive Extragenic Palindromic (REP) sequence has been used to identify environmental bacterial species and strains. We applied REP-PCR to detect and differentiate four major Gram-negative food-borne bacterial pathogens, E. coli, Salmonella, Shigella, and Vibrio. Target DNA fragments of these pathogens were amplified by REP-PCR method. PCR-generated DNA fragments were separated on 1.5% agarose gel. Dendrograms for PCR products of each strain were constructed using photo-documentation system. REP-PCR reactions with primer pairs REP1R-I and REP2-I revealed distinct REP-PCR-derived genomic fingerprinting patterns from E. coli, Salmonella, Shigella, and Vibrio. REP-PCR method provided clear distinctions among different bacterial species containing REP-repetitive elements and can be widely used for typing food-borne Gram-negative strains. Results showed established REP-PCR reaction conditions and generated dendrograms could be used with other supplementary genotyping or phenotyping methods to identify isolates from outbreak and to estimate relative degrees of genetic similarities among isolates from different outbreaks to determine whether they are clonally related.
Keywords
foodborne bacterial pathogens; REP-PCR; genotyping; genetic similarity;
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