Korean Journal of Food Science and Technology (한국식품과학회지)

Korean Society of Food Science and Technology (한국식품과학회)
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Development of Immunochromatography for Detection of Escherichia coli O157:H7 Contaminated in Meats and Sprouts

육류 및 새싹채소에 오염된 Escherichia coli O157:H7의 신속검출을 위한 면역크로마토그래피법의 개발

  • Lee, Won-Gyeong (Division of Applied Life Science (BK21 program), Gyeongsang National University) ;
  • Ryu, Hee-Jeong (Division of Applied Life Science (BK21 program), Gyeongsang National University) ;
  • Song, Jeong-Un (Division of Applied Life Science (BK21 program), Gyeongsang National University) ;
  • Kim, Kyeong-Yeol (Division of Applied Life Science (BK21 program), Gyeongsang National University) ;
  • Chung, Do-Yeong (Department of Food Science & Technology, Gyeongsang National University) ;
  • Kim, Jeong-Sook (Institute of Agriculture and Life Science, Gyeongsang National University) ;
  • Shim, Won-Bo (Institute of Agriculture and Life Science, Gyeongsang National University) ;
  • Chung, Duck-Hwa (Division of Applied Life Science (BK21 program), Gyeongsang National University)
  • 이원경 (경상대학교 응용생명과학부(BK21 program)) ;
  • 류희정 (경상대학교 응용생명과학부(BK21 program)) ;
  • 송정언 (경상대학교 응용생명과학부(BK21 program)) ;
  • 김경열 (경상대학교 응용생명과학부(BK21 program)) ;
  • 정도영 (경상대학교 식품공학과) ;
  • 김정숙 (경상대학교 농업생명과학연구원) ;
  • 심원보 (경상대학교 농업생명과학연구원) ;
  • 정덕화 (경상대학교 응용생명과학부(BK21 program))
  • Received : 2011.04.21
  • Accepted : 2011.06.29
  • Published : 2011.10.31
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Abstract

Escherichia coli O157:H7 has been considered as a significant food-borne pathogen since its role in causing hemorrhagic colitis and hemolytic uremic syndrome in humans was recognized. In this study, we developed an immunochromatography (ICG) assay for the detection of E. coli O157:H7. E. coli O157:H7 monoclonal antibody (EC MAb) and colloidal gold were conjugated and its specificity was determined by the ICG treated with EC MAb and antimouse IgG at test and control lines, respectively. The detection limit of the ICG was $1{\times}10^5$ CFU/mL, and no crossreactivity was observed to other E. coli strains and major food-borne pathogens. To determine the minimum enrichment time for the ICG, meats and sprouts were inoculated with $1{\times}10$ CFU/100 ${\mu}L$ of E. coli O157:H7. After enrichment time of 10 and 2 h for meats and sprouts, respectively, up to $1{\times}10$ CFU/100 ${\mu}L$ of E. coli O157:H7 could be detected by ICG.

E. coli O157:H7은 요독증후군, 출혈성 대장염 및 설사 등을 유발하는 원인균으로, 전세계적으로 이들의 오염에 따른 식중독 발생이 증가하고 있어, 본 연구에서는 E. coli O157:H7을 신속 정확하게 분석할 수 있는 면역크로마토그래피법을 개발하였고, 보다 신속한 검출을 위하여 육류와 새싹시료를 대상으로 최소 증균시간을 확인하였다. 먼저, 면역크로마토그래피법의 개발을 위해 colloidal gold와 EC MAb를 합성하여 EC MAb-gold conjugate를 제작한 다음, 접합여부를 확인한 결과, test line과 control line에 각각 처리되는 EC MAb와 anti-mouse IgG에 특이적으로 반응하였다. 제작된 EC MAb-gold conjugate를 이용하여 개발된 면역크로마토그래피법의 검출한계는 $1{\times}10^5$ CFU/mL 수준까지 검출이 가능한 것으로 확인되었고, 다른 대장균속 및 주요 식중독균과의 교차반응성은 나타나지 않았다. 개발된 면역크로마토그래피법을 이용하여 임의로 E. coli O157:H7을 오염시킨 돼지고기, 소고기, 닭고기 및 새싹채소를 증균하는 동안 분석한 결과, 돼지고기와 닭고기는 6시간, 소고기는 10시간, 특히 새싹채소는 2시간 후부터 $1{\times}10$ CFU/100 ${\mu}L$ 수준까지 검출이 가능한 것으로 나타났다. 따라서 본 연구에서 개발된 면역크로마토그래피법을 이용하여 육류와 채소 등에 오염된 E. coli O157:H7을 최대 10시간 이내로 신속히 분석할 수 있을 것으로 판단된다.

Keywords

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