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


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.


  1. Farber JM, Johnston MA, Purvis D, Loit A. Surveillace of soil and semi-soft cheese or the presence of Listeria spp. Int. J. Food Microbiol. 5: 157-163 (1987)
  2. Farber JM, Peterkin PI. Listeria monocytogenes, a food-borne pathogen. Microbiol. Rev. 55: 476-511 (1991)
  3. Hahm BK, Bhunia AK. Effect of environmental stresses on antibody-based detection of Escherichia coli O157:H7, Salmonella enterica serotype Enteritidis and Listeria monocytogenes. J. Appl. Microbiol. 100: 1017-1027 (2006)
  4. Bahk JR, Marth EH. Listeriosis and Listeria monocytogenes. Korean J. Appl. Microbiol. Bioeng. 17: 634-644 (1989)
  5. Frens G. Preparation of gold dispersions of varying particle size: controlled nucleation for the regulation of the particle size in monodisperse gold suspensions. Nature-Phys. Sci. 241: 20-22 (1973)
  6. Jung BY, Jung SC. Development of the rapid detection kit for Salmonella spp. using immunochromatographic assay. Korean J. Vet. Res. 45: 191-197 (2005)
  7. Sewell AM, Warburton DW, Boville A, Daley EF, Mullen K. The development of an efficient and rapid enzyme linked fluorescent assay method for the detection of Listeria spp. from foods. Int. J. Food Microbiol. 81: 123-129 (2003)
  8. Chiao DJ, Shyu RH, Hu CS, Chiang HY, Tang SS. Colloidal gold-based immunochromatographic assay for detection of botulinum neurotoxin type B. J. Chromatogr. B 809: 37-41 (2004)
  9. Kerdahi KF, Istafanos PF. Rapid determination of Listeria monocytogenes by automated enzyme-linked immunoassay and nonradioactive DNA probe. J. Assoc. Off. Ana. Chem. Int. 83: 86-88 (2000)
  10. Mckearn TJ, Weiss A, Stuart FP, Fitch FW. Selective suppression of humoral and cell-mediated immune responses to rat alloantigens by monoclonal antibodies produced by hybridoma cell lines. Transplant. P. 11: 932-935 (1979)
  11. Geoghegan WD, Ackerman GA. Adsorption of horseradish peroxidase, ovomucoid and anti-immunoglobulin to colloidal gold for the indirect detection of concanavalin A, wheat germ agglutinin and goat anti-human immunoglobulin G on cell surfaces at the electron microscopic level: a new method, theory and application. J. Histochem. Cytochem. 25: 1187-1200 (1977)
  12. Fleming DW, Cochi SL, MacDonald KL, Brondum J, Hayes PS, Plikaytis BD, Holmes MB, Audurier A, Broome CV, Reingold AL. Pasteurized milks a vehicle of infection in an outbreak of listeriosis. New Engl. J. Med. 312: 404-407 (1985)
  13. Kohler G, Milstein C. Continuous cultures of fused cells producing antibody of predefined specificity. Nature 256: 495-501 (1975)
  14. Jung BY, Jung SC, Kim JM. Development of the immunochromatographic strip for the rapid detection of Listeria spp. Korean J. Vet. Res. 45: 169-177 (2005)
  15. Paek SH, Lee SH, Cho JH, Kim YS. Development of rapid one-step immunochromatographic assay. Methods 22: 53-60 (2000)
  16. Siragusa GR and Johnson MG. Monoclonal antibody specific for listeria monocytogenes, Listeria innocua, Listeria welshimeri. Appl. Environ. Microb. 56: 1897-1904 (1990)
  17. Kim SH, Park MK, Kim JY, Chuong PD, Lee YS, Yoon BS, Hwang KK, Lim YK Development of a sandwich ELISA for the detection of Listeria spp. using specific flagella antibodies. J. Vet. Sci. 6: 41-46 (2005)