Korean Journal of Food Science and Technology (한국식품과학회지)

Korean Society of Food Science and Technology (한국식품과학회)
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Using Reverse Dot Hybridization Method and 16S rRNA Gene (16S rDNA) for Identifying the Food Poisoning Microorganism in Foods

Reverse dot hybridization 방법과 16S rRNA gene(16S rDNA)을 이용한 식품에서 식중독균의 탐색

  • 김민성 (한양대학교 자연과학대학 생명과학과) ;
  • 신규철 (한양대학교 자연과학대학 생명과학과) ;
  • 이형구 (한양대학교 자연과학대학 생명과학과) ;
  • 한명수 (국가지정 물환경생태복원연구실) ;
  • 민병례 (상명대학교 자연과학대학 생명과학과) ;
  • 최영길 (한양대학교 자연과학대학 생명과학과)
  • Published : 2003.06.01
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Abstract

DNA sequence information on small-subunit rRNA gene (16S rDNA) obtained from food-poisoning bacterial culture was used to investigate the presence of bacterial pathogens in food. By reverse dot blot detection method, presence of food-poisoning bacteria could be confirmed on hybridization of digoxigenin-labeled 16S rDNA Polymerase Chain Reaction (PCR) primer product and biotin-labeled specific oligonucleotide probe. Escherichia coli, Bacillus cereus. and Salmonella sp. were used as the representative food-poisoning bacterial microorganisms. An oligonucleotide probe, based on the variable region of 16S rRNA gene, was used as the specific probe. These tools may be more useful than classic biochemical method for rapid identification of contaminated food.

식중독은 세균에 의한 발병이 대부분이다. 따라서 식품에서 식중독 원인균을 신속하게 탐색하게 식중독으로부터의 되면 피해를 줄일 수 있을 것이다. 고전적인 식중독 원인균 탐색은 증균, 선택적 배지를 이용한 isolation, 생화학적 특징을 활용하는 분석이 있으나 많은 시간이 소요되는 단점을 갖고 있었다. 본 연구는 16S rRNA gene(16S rDNA)로부터 얻은 DNA 염기 서열을 이용 식중독 원인균의 특이적 oligonucleotide probe을 제작 reverse dot blot hybridization과 PCR 방법을 이용하여 고전적인 방법보다 빠른 시간 내에 식품에서 원인균을 탐색 할 수 있었다. 우유를 인공적으로 본 연구에서 사용한 균주로 오염시킨 후 DNA를 추출하여 PCR 증폭산물과 oligonucleotide probe를 hybridization 시킨 결과 oligonucleotide probe를 hybridization 시킨 결과 oligonucleotide probe가 위치한 곳에서 발색 반응이 나타났다. 본 연구에서 본 연구를 통해 DNA microchip으로 활용 짧은 시간 내에 많은 종류의 식중독 원인균을 탐색 할 수 있는 가능성을 확인하였다.

Keywords

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