Production of Monoclonal Antibody against Escherichia coli O157:H7 and Development of Enzyme Linked Immunosorbent Assay

Escherichia coli O157:H7의 단크론성 항체 생산과 효소면역분석법의 개발

  • Ryu, Hee-Jeong (Division of Applied Life Science (BK21 program), Gyeongsang National University) ;
  • Kim, Jeong-Sook (Institute of Agriculture and Life Science, Gyeongsang National University) ;
  • Kim, Kyeong-Yeol (Division of Applied Life Science (BK21 program), Gyeongsang National University) ;
  • Nam, Bo-Ram (Division of Applied Life Science (BK21 program), Gyeongsang National University) ;
  • Nam, Min-Ji (Division of Applied Life Science (BK21 program), Gyeongsang National University) ;
  • Shim, Won-Bo (Institute of Agriculture and Life Science, Gyeongsang National University) ;
  • Kim, Nam-Soo (Food Bio-nano Technology Research Group, Korea Food Research Institute) ;
  • Cho, Yong-Jin (Food Bio-nano Technology Research Group, Korea Food Research Institute) ;
  • Chung, Duck-Hwa (Division of Applied Life Science (BK21 program), Gyeongsang National University)
  • 류희정 (경상대학교 응용생명과학부(BK 21 program)) ;
  • 김정숙 (경상대학교 농업생명과학연구원) ;
  • 김경열 (경상대학교 응용생명과학부(BK 21 program)) ;
  • 남보람 (경상대학교 응용생명과학부(BK 21 program)) ;
  • 남민지 (경상대학교 응용생명과학부(BK 21 program)) ;
  • 심원보 (경상대학교 농업생명과학연구원) ;
  • 김남수 (한국식품연구원 바이오나노연구단) ;
  • 조용진 (한국식품연구원 바이오나노연구단) ;
  • 정덕화 (경상대학교 응용생명과학부(BK 21 program))
  • Received : 2010.01.11
  • Accepted : 2010.02.10
  • Published : 2010.06.30

Abstract

Escherichia coli O157:H7 causes hemolytic uremic syndrome and hemorrhagic colitis in humans. The objectives of this study were to produce monoclonal antibody(MAb) against E. coli O157:H7 and to develop an enzyme linked immunosorbent assay(ELISA) for the rapid detection of E. coli O157:H7 in agri-stockbreeding. The characterization of MAb produced from hybridoma cell (HKEC 4G8-5) was validated by ELISA and Western blot. The produced MAb was specific to E. coli O157:H7 and showed weak cross-reaction to Staphylococcus aureus. The detection limit of ELISA based on 4G8-5 MAb was $10^5\;cell/mL$. Although the ELISA could not detect E. coli O157:H7 in the meat and sprout samples inoculated with $1{\times}10^1\;cell$/10 g without enrichment, the same samples after enrichment for 6 hr were confirmed to be positive by ELISA. These results indicated that the ELISA combined with short enrichment (6 hr) is useful tool for rapid screening of E. coli O157:H7 in various samples.

Keywords

E. coli O157:H7;monoclonal antibody;ELISA

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