Korean Journal of Food Science and Technology (한국식품과학회지)

Korean Society of Food Science and Technology (한국식품과학회)
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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
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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.

본 연구에서는 최근 식품이나 농축산물 등에서 빈번하게 검출되고 있는 장관출혈성 대장균인 E. coli O157:H7에 대한 단크론성 항체를 생산하고, 신속 정확하게 검색할 수 있는 간접효소면역분석법을 개발한 후 시료적용가능성을 확인하였다. 먼저 LPS, HKEC 및 FKEC를 E. coli O157:H7의 면역원으로 준비하여 mouse 면역에 사용하였고, 세포융합과 cloning을 실시하여 단클론성 항체를 생산하였으며, 생산된 항체 중 높은 역가를 나타낸 HKEC 4G8-5 항체로 ID-ELISA법을 개발하였다. 개발된 IDELISA법의 최저검출한계는 $10^5\;cell/mL$이었고, S. aureus와 Sal. Typhimurium에 대해서 약간의 반응성을 나타내었지만 E. coli O157:H7에 대해 보다 강한 반응성을 나타내었기 때문에 E. coli O157:H7에 매우 특이적인 분석법인 것으로 확인되었다. 개발된 ID-ELISA법을 이용하여 돼지고기, 소고기, 닭고기 및 새싹채소에 대한 검출률 향상을 위해 각 시료를 증균한 결과, 3종의 육류와 새싹채소는 각각 증균 6시간과 2시간부터 검출이 가능한 것으로 나타나 기존의 증균법보다 신속하게 시료 속 E. coli O157:H7을 분석할 수 있을 것으로 판단된다. 따라서 본 연구에서 개발된 IDELISA법을 적용하여 농축산물 중에 존재하는 E. coli O157:H7을 분석한다면 보다 신속하고 간편하게 분석할 수 있을 것으로 판단된다.

Keywords

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