동아시아식생활학회지 (Journal of the East Asian Society of Dietary Life)

  • 제19권2호
  • /
  • Pages.295-300
  • /
  • 2009
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  • 1225-6781(pISSN)
  • /
  • 2288-8802(eISSN)
동아시아식생활학회 (The East Asian Society of Dietary Life)
권호 표지

PCR을 이용한 육류 내 Salmonella sp. 및 Salmonella Typhimurium 분리 검출

Selective Detection of Salmonella sp. and Salmonella Typhimurium in Meat by Polymerase Chain Reaction

  • 주종원 (중부대학교 애완동물자원학과) ;
  • 홍경표 (중부대학교 호텔외식산업학과) ;
  • 김용휘 (중부대학교 호텔외식산업학과) ;
  • 조상범 (건국대학교 동물자원연구센터)
  • Joo, Jong-Won (Dept. of Companion Animal and Animal Resources Science, Joongbu University) ;
  • Hong, Kyung-Pyo (Dept. of Hotel and Foodservice Industry, Joongbu University) ;
  • Kim, Yong-Hui (Dept. of Hotel and Foodservice Industry, Joongbu University) ;
  • Cho, Sang-Buem (Animal Resources Research Center, Konkuk University)
  • 투고 : 2009.02.26
  • 심사 : 2009.03.26
  • 발행 : 2009.04.30
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초록

본 연구에서는 육류 식품 시료에서 단시간 내에 살모넬라를 검출하기 위하여 PCR을 이용한 검출용 프라이머들의 특이성과 민감성을 평가하였다. 실험에 사용된 프라이머들은 Salmonella Typhimurium의 mdh와 invA 유전자의 염기서열에 기초하여 제작되었다. 각각의 primer들의 검출 감도를 평가하여 최종적으로 broad spectrum primer SLM1과 S. Typhimurium specific primer SLT4를 선발하였다. 또한, 시료에 오염된 병원균의 최소 검출량이 어느 정도인지를 확인하기 위하여 살모넬라 균수를 reaction tube당 $10^0{\sim}10^3$ cell까지 다양하게 하여 검출 감도를 측정한 결과, 최소 1 cell에서도 PCR 산물을 나타내어 검출 감도가 매우 우수한 것으로 나타났다. 살모넬라 균주를 혼합한 소고기와 돼지고기 시료에서 각각 프라이머들의 검출 감도를 평가한 결과, 증균하지 않은 시료 자체와 세균학적 방법으로 증균 배양한 시료 그리고 증균 배양된 시료로부터 세균의 DNA를 추출한 시료 간에 큰 차이를 나타내지 않았다. 이에 육류 식품에서 살모넬라 혹은 S. Typhimurium의 검출을 위한 프라이머로서 효율적으로 사용이 가능할 것으로 판단된다.

The specificity and sensitivity of oligonucleotide primers were examined for the rapid detection of Salmonella in meat samples. The oligonucleotide primers used in this study were designed with the modification of mdh and invA sequence in the chromosome of Salmonella Typhimurium. Through the subsequent analysis of the specificity and sensitivity of the primers, two types of oligonucleotide primers, SLM1 and SLT4 were selected for the detection of Salmonella genus specific and S. Typhimurium species specific, respectively. The lowest detection limit of each primer was represented as 1 cell per reaction when reacted with a prepared DNA solution. The detection efficiency of the two primers was analysed with beef and pork samples intentionally contaminated with a mixture of Salmonella culture, and three preparation methods -, namely direct reaction after extraction, enrichment after extraction, and DNA extraction after enrichment for PCR reaction, - were also compared. No differences were found in the results according to meat sources and preparation methods.

키워드

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