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Establishment of Sample Preparation Method to Enhance Recovery of Food-borne Pathogens from Produce

농산물 중 식중독세균 검출을 위한 전처리법 확립

  • 김세리 (농촌진흥청 국립농업과학원 농산물안전성부 유해생물팀) ;
  • 최송이 (농촌진흥청 국립농업과학원 농산물안전성부 유해생물팀) ;
  • 서민경 (농촌진흥청 국립농업과학원 농산물안전성부 유해생물팀) ;
  • 이지영 (경인지방식품의약품안전청 수입관리과) ;
  • 김원일 (농촌진흥청 국립농업과학원 농산물안전성부 유해생물팀) ;
  • 윤요한 (숙명여자대학교 식품영양학과) ;
  • 류경열 (농촌진흥청 연구정책국 연구성과관리과) ;
  • 윤종철 (농촌진흥청 국립농업과학원 농산물안전성부 유해생물팀) ;
  • 김병석 (농촌진흥청 국립농업과학원 기획조정과)
  • Received : 2013.07.01
  • Accepted : 2013.09.12
  • Published : 2013.09.30

Abstract

To establish sample preparation method for detection of food-borne pathogens from lettuce, perilla leaves, cucumber, pepper, and cherry tomato, the influences of diluent composition, processing time, and proportion of diluent to sample were examined. Each produce was inoculated with 6.0 log $CFU/cm^2$ of Escherichia coli O157:H7, Salmonella Typhimurium, Listeria monocytogenes, Staphylococcus aureus, and Bacillus cereus. Each produce was treated with 0.1% peptone water, and D/E neutralizing broth. Processing time of produce was 30, 60, 90, and 120s, and the proportion of diluent to sample was 2 : 1, 4 : 1, 9 : 1, and 19 : 1. The number of bacteria after treatment of D/E neutralizing broth was higher than that of 0.1% peptone water (P<0.05). In cherry tomato, the population of S. typhimurium recovered from treated with D/E broth was higher than that recovered from treated with 0.1% peptone water by 1.05 log $CFU/cm^2$ (P<0.05). No difference in numbers of pathogens was observed in processing time. Optimum proportion of diluent to perilla leaf, iceberg lettuce, cucumber, green pepper, and tomato was 9 : 1, 4 : 1, 2 : 1, 2 : 1, and 2 : 1, respectively. These data suggest that D/E neutralizing broth should be recommend as diluent, and the diluent volume applied to produce should be determined in proportion to produce surface area per weight (g).

본 연구는 농산물 중 식중독세균 검출을 위한 시료 전처리법을 확립하기 위하여 전처리용액, 전처리 시간, 전처리용액과 시료의 비율에 따른 회수율을 비교하였다. 이를 위하여 양상추, 들깻잎, 오이, 고추, 방울토마토에 E. coli O157:H7, S. Typhimurium, L. monocytogenes, S. aureus, B. cereus를 6.0 log $CFU/cm^2$로 접종하고 0.1% 펩톤수와 D/E broth로 처리하였다. 또한 처리시간은 30, 60, 90, 120초, 시료와 전처리용액의 비율은 1 : 2, 1 : 4, 1 : 9, 1 : 19로 각각 처리하였다. 그 결과, D/E broth로 처리하였을 때 0.1% 펩톤수로 처리하였을 때에 비하여 전반적으로 식중독균의 회수율이 높았다(P<0.05). 특히 방울토마토의 경우, D/E broth로 처리한 구의 S. Typhimurium의 수는 0.1% 펩톤수로 처리한 구 보다 1.05 log $CFU/cm^2$ 높았다(P<0.05). 또한 들깻잎, 양상추, 오이, 고추, 토마토와 전처리용액의 최적 비율은 1 : 9, 1 : 4, 1 : 2, 1 : 2, 1 : 2였다. 한편, 처리시간에 따른 회수율의 유의적인 차이는 없었다. 따라서 본 연구 결과를 통해 볼 때 농산물 중 식중독균 검출을 위한 전처리용액은 D/E broth가 적절한 것으로 판단되며 표면적에 따라 시료와 전처리 용액의 비율을 결정하는 것이 분석의 정확성을 높일 수 있을 것으로 사료된다.

Keywords

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