Korean Journal of Food Science and Technology (한국식품과학회지)

Korean Society of Food Science and Technology (한국식품과학회)
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Development of Immunochromatography for the Rapid Detection of Listeria monocytogenes

Listeria monocytogenes 신속 검출을 위한 면역크로마토그래피법의 개발

  • Choi, Jin-Gil (Division of Applied Life Science(BK21), Graduate School of Gyeongsang National University) ;
  • Shim, Won-Bo (Division of Applied Life Science(BK21), Graduate School of Gyeongsang National University) ;
  • Je, Jung-Hyun (Division of Applied Life Science(BK21), Graduate School of Gyeongsang National University) ;
  • Kim, Ji-Young (Division of Applied Life Science(BK21), Graduate School of Gyeongsang National University) ;
  • Lee, Kyu-Ho (Environmental Science, HanKuk University of Foreign Studies) ;
  • Kim, Min-Gon (Korea Research Institute of Bioscience and Biotechnology) ;
  • Ha, Sang-Do (Department of Food Science & Technology, Chung-Ang University) ;
  • Kim, Keun-Sung (Department of Food Science & Technology, Chung-Ang University) ;
  • Kim, Kwang-Yup (Department of Food Science & Technology, Chung-Ang University) ;
  • Kim, Cheol-Ho (Department of Biological Sciences, Sungkyunkwan University) ;
  • Chung, Duck-Hwa (Division of Applied Life Science(BK21), Graduate School of Gyeongsang National University)
  • 최진길 (경상대학교 대학원 응용생명과학부(BK21)) ;
  • 심원보 (경상대학교 대학원 응용생명과학부(BK21)) ;
  • 제정현 (경상대학교 대학원 응용생명과학부(BK21)) ;
  • 김지영 (경상대학교 대학원 응용생명과학부(BK21)) ;
  • 이규호 (한국외국어대학교 환경학과) ;
  • 김민곤 (한국생명공학연구원) ;
  • 하상도 (중앙대학교 식품공학과) ;
  • 김근성 (중앙대학교 식품공학과) ;
  • 김광엽 (충북대학교 식품공학과) ;
  • 김철호 (성균관대학교 생명과학과) ;
  • 정덕화 (경상대학교 대학원 응용생명과학부(BK21))
  • Published : 2007.06.30
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Abstract

The objective of this study was the development of immunochromatography (ICG) for the rapid and accurate detection of Listeria monocytogenes. Here, monoclonal antibodies (MAb) were conjugated with 40 nm colloidal gold particles, where the conjugate was used as the detection reagent in the ICG. The ICG was composed of three pads (sample, conjugate, and absorbance pads) and one nitrocellulose membrane. The colloidal gold-MAb conjugate was applied to the conjugate pad, and the test line and control line on the membrane were treated with MAb (FKLM-3BI2-37) and anti-mouse IgG, respectively. The detection limit of the ICG was $10^{5}$ cell/mL and it showed no cross-reaction to food borne pathogens. We inoculated meat and lettuce samples with various counts of L. monocytogenes, and analyzed them by ICG. All the inoculated meat samples gave positive results after enrichment for 24 h in LEB. These results indicate that ICG was able to serve as a primary screening tool for L. monocytogenes in various foods and agricultural products within 20 min after enrichment.

본 연구에서는 L. monocytogenes를 신속하게 분석할 수 있는 면역크로마토그래피법을 개발하고자 하였다. 먼저 L. monocytogenes에 대한 단크론성 항체(FKLM-3B12-37)를 개발하여 직경 40nm로 제작된 colloidal gold와 합성한 후 conjugate pad에 처리하였고 nitrocellulose membrane 상의 test line과 control line에 단크론성 항체와 anti-mouse IgG를 각각 처리하여 면역크로마토그래피법을 개발하였다. 개발된 면역크로마토그래피법의 검출한계는 $10^{5}$ cell/mL 수준까지 검출이 가능했으며 다른 식중독 세균과는 교차 반응이 없는 것으로 나타났다. 임으로 오염시킨 식품시료의 경우 증균과정 없이 $10^{4}$ cell/25g까지 바로 분석하는데 어려움이 있으며 24시간의 증균과정을 거친 후에는 $10^{4}$ cell/25g까지 검출할 수 있었다. 본 연구에서 개발된 면역크로마토그래피법은 24시간의 증균 만으로 식품에 오염되어있는 L. monocytogenes를 검출할 수 있는 것으로 사료되었고, 기존의 식중독균 진단방법에 비해 신속하고 간편하게 그 결과를 확인할 수 있었다.

Keywords

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