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유제품 및 가공식품에서 Listeria monocytogenes 검출을 위한 배지법과 신속 검사키트의 유효성 검증

Evaluation of Conventional Culture Methods and Validation of Immunoassays for Rapid Detection of Listeria monocytogenes in Dairy and Processed Foods

  • 한소리 (건국대학교 수의과대학 공중보건학) ;
  • 현지연 (건국대학교 수의과대학 공중보건학) ;
  • 김희연 (서울지방식품의약품안전청 시험분석과) ;
  • 박종석 (식품의약품안전청 연구기획조정관) ;
  • 허석 (식품의약품안전청 연구기획조정관) ;
  • 신호철 (건국대학교 수의과대학 수의약리독성학) ;
  • 서건호 (건국대학교 수의과대학 공중보건학)
  • Han, So-Ri (Department of Public Health, College of Veterinary Medicine, Konkuk University) ;
  • Hyeon, Ji-Yeon (Department of Public Health, College of Veterinary Medicine, Konkuk University) ;
  • Kim, Hee-Yun (Testing & Analysis Division, Seoul Regional Korea Food & Drug Administration) ;
  • Park, Jong-Seok (Research Planning Management Office, Korea Food & Drug Administration) ;
  • Heo, Seok (Research Planning Management Office, Korea Food & Drug Administration) ;
  • Shin, Ho-Chul (Department of Veterinary Pharmacology and Toxicology, College of Veterinary Medicine, Konkuk University) ;
  • Seo, Kun-Ho (Department of Public Health, College of Veterinary Medicine, Konkuk University)
  • 발행 : 2008.12.31

초록

L. monocytogenes는 최근 전 세계적으로 주목받고 있는 식중독의 원인체 중 하나로서 면역력이 약한 사람에게서 심각한 리스테리아증을 유발한다. 이 때문에 안전한 식품제공과 공중보건 증진 측면에서 L. monocytogenes균을 검출하는 것은 매우 중요하다. 본 연구의 목적은 현재 사용되고 있는 배지법에서 배양시간에 따른 검출능력을 비교해보고, 상용화된 L. monocytogenes 신속검출 키트의 유효성을 검증하는 것이다. 다양한 식품 샘플(훈제연어, 생식용 두부, 우유, 샐러드)에 총 20개의 샘플에서 일부분의 양성 결과가 나오도록 L. monocytogenes를 접종하였다. 이를 예비 증균과 배지에서의 배양을 거쳐 마지막으로 $Microgen^{TM}$ Listeria-ID를 이용하여 균을 확인하였다. 신속검출 키트인 $VIDAS^{(R)}$ Listeria monocytogenes II(LMO2)와 $REVEAL^{(R)}$ for Listeria 도 시행하여 배지법과 검출능력을 비교하였다. 배지법에서 배양시간을 24시간에서 48시간으로 늘리면 훈제연어는 0개에서 9개,두부는 0개에서 10개로, 우유는 2개에서 17개로, 샐러드는 4개에서 16개로 모든 식품 샘플에서 양성결과가 눈에 띄게 증가하였다. 따라서 24시간을 채택하고 있는 배지법에서의 예비증균 시간을 48시간으로 늘리는 것이 필요하다 할 수 있겠다. 신속검출키트와 배지법 사이에는 통계학적 유의차가 없는 것으로 나타나 배지법을 시행하기에 앞서 예비양성시험(presumptive screening)으로 활용하기에 알맞다고 사료된다.

Listeria monocytogenes is a foodborne pathogen inducing listeriosis in human. We compared two different culture methods for detection of L. monocytogenes and validated two commercial kits, $VIDAS^{(R)}$ and $REVEAL^{(R)}$ for Listeria. L. monocytogenes was inoculated into various food samples to generate partial positive samples. The inoculated samples were enriched in half-Fraser broth for 48 hr at $30^{\circ}C$. The enriched samples were streaked onto Oxford agar at 24 and 48 hr postincubation followed by biochemical confirmation and concurrently analyzed by using the two commercial kits for comparison. When the enrichment period was extended from 24 to 48 hr, the numbers of positive samples were dramatically increased from 6 to 52 out of 80 samples tested using the culture method. With the commercial kits, the numbers of positive samples were also significantly increased from 10 to 18 and 1 to 18, respectively, when the enrichment period was extended from 48 to 72 hr. There was no statistical difference between the 24 hr culture method and $VIDAS^{(R)}$ or $Reveal^{(R)}$ with 48 hr enrichment. In conclusion, the 24 hr for the culture method was insufficient to detect L. monocytogenes in various foods. The commercial kits could be adequate means for presumptive screening of L. monocytogenes in food.

키워드

참고문헌

  1. Barocci, S., Calza, L., Blasi, G., Briscolini, S., Curtis, M. D., Palombo, B., Cucco, L., Postacchini, M., Sabbatini, M., Graziosi, T., Nardi, S., and Pezzotti, G. (2008) Evaluation of a rapid molecular method for detection of Listeria monocytogenes directly from enrichment broth media. Food Control 19, 750-756 https://doi.org/10.1016/j.foodcont.2007.07.013
  2. Beumer, R. R., te Giffel, M. C., and Rombouts, F. M. (1997) A comparison of rapid methods for the detection of Listeria spp. and Listeria monocytogenes. In: Beumer, R. (ed), Listeria monocytogenes detection and behaviour in food and in the envirionment (Thesis Landbouwuniversiteit Wageningen), Koninklijke Bibliotheek, Den Haag, Netherlands, pp. 69-86
  3. Chapman, P. A. and Ashton, R. (2003) An evaluation of rapid methods for detecting Escherichia coli O157 on beef carcasses. Int. J. Food Microbiol. 87, 279-285 https://doi.org/10.1016/S0168-1605(03)00074-6
  4. Choi, J. G., Shim, W. B., Je, J. H., Kim, J. Y., Lee, H. H., Kim, M. G., Ha, S. D., Kim, K. S., Kim, K. Y., Kim, C. H., and Chung, D. H. (2007) Development of immunochromatography for the rapid detection of Listeria monocytogenes. Korean J. Food Scl.Technol. 39, pp. 299-303
  5. Fahey, J. W., Ourisson, P. J., and Degnan, F. H. (2006) Pathogen detection, testing, and control in fresh broccoli sprouts. Nutr. J. 21, 5-13
  6. Kim, S. H., Kim, J. Y., Han, W., Jung, B. Y., Chuong, P. D., Joo, H. J., Ba, H. V., Son, W. G., Jee, Y. H., Yoon, B. S., Lee, Y. S., and Lim, Y. K. (2007) Development and evaluation of an immunochromatographic assay for screening Listeria spp. in pork and milk. Food Sci. Biotechnol. 16, 515-519
  7. Kim, Y. S. and Ha, S. D. (2006) Detection and isolation of foodborne bacteria using conventional culture media. Safe Food 1, 5-15
  8. Mena, C., Almeida, G., Carneiro, L., Teixeira, P., Hogg, T., and Gibbs, P. A. (2004) Incidence of Listeria monocytogenes in different food products commercialized in Portugal. Food Microbiol. 21, 213-216 https://doi.org/10.1016/S0740-0020(03)00057-1
  9. Midelet-Bourdin, G., Leleu, G., and Malle, P. (2007) Evaluation of the international reference methods NF EN ISO 11290-1 and 11290-2 and an in-house method for the isolation of Listeria monocytogenes from retail seafood products in france. J Food Prot. 70, 891-900 https://doi.org/10.4315/0362-028X-70.4.891
  10. O' Grady, J., Sedano-Balbas, S., Maher, M., Smith, T., and Barry, T. (2008) Rapid real-time PCR detection of Listeria monocytogenes in enriched food samples based on the ssrA gene, a novel diagnostic target. Food Microbiol. 25, 75-84 https://doi.org/10.1016/j.fm.2007.07.007
  11. Oravcova, K., Trncikova, T., Kuchta, T., and Kaclikova, E. (2008) Limitation in the detection of Listeria monocytogenes in food in the presence of competing Listeria innocua. J Appl. Microbiol. 104, 429-437
  12. Ramaswamy, V., Cresence, V. M., Rejitha, J. S., Lekshmi, M. U., Dharsana, K. S., Prasad, S. P., and Vijila, H. M. (2007) Listeria-review of epidemiology and pathogenesis. J Microbiol. Immunol. Infect. 40, 4-13
  13. Seo, K. H., Jeong, D. H., Kim, H. Y., and Jeong, D. G. (2006) Guide line and effective validation of rapid kits for food hazardous materials. KFDA 2006 reserch report, http://rnd.kfda. go.kr/index.jsp
  14. Vaz-Velho, M., Duarte, G., and Gibbs, P. (2000) Evaluation of mini-VIDAS rapid test for detection of Listeria monocytogenes from production lines of fresh to cold-smoked fish. J Microbiol. Methods 40, 147-151
  15. http://www.aoac.org/
  16. 김명숙, 김옥미, 김인구, 이경임, 이선희, 이정갑, 임성미, 정성태, 조좌형 (2002) 식품위생학. 훈민사, 서울

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