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

주요 식중독 그람 음성 세균 4속의 REP-PCR genotyping

  • 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)
  • 정혜진 (중앙대학교 식품공학과) ;
  • 서현아 (중앙대학교 식품공학과) ;
  • 김영준 (중앙대학교 식품공학과) ;
  • 조준일 (중앙대학교 식품공학과) ;
  • 김근성 (중앙대학교 식품공학과)
  • Published : 2005.08.31

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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