Journal of the Korean Society of Food Science and Nutrition (한국식품영양과학회지)

The Korean Society of Food Science and Nutrition (한국식품영양과학회)
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Detection of Clostridium difficile by Loop-Mediated Isothermal Amplification

등온증폭법을 이용한 Clostridium difficile 검출

  • In, Ye-Won (Dept. of Food and Nutrition, Kookmin University) ;
  • Ha, Su-Jeong (Dept. of Food and Nutrition, Kookmin University) ;
  • Yang, Seung-Kuk (Dept. of Food and Nutrition, Kookmin University) ;
  • Oh, Se-Wook (Dept. of Food and Nutrition, Kookmin University)
  • 인예원 (국민대학교 식품영양학과) ;
  • 하수정 (국민대학교 식품영양학과) ;
  • 양승국 (국민대학교 식품영양학과) ;
  • 오세욱 (국민대학교 식품영양학과)
  • Received : 2012.05.18
  • Accepted : 2012.07.10
  • Published : 2012.09.30
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Abstract

This study was conducted to develop a loop-mediated isothermal amplification (LAMP) method for the detection of Clostridium difficile. The tested target gene was 16S ribosomal RNA. Five different LAMP primer sets were designed, and LAMP was performed. All primer sets targeting the 16S rRNA gene (BIP, FIP, B3, F3, LF, PF) were determined as positive in tcdA-positive, tcdB-postive ($A^+B^+$) and tcdA-negative, tcdB-negative ($A^-B^-$) Clostridium difficile strains. As the LAMP reaction took less than 80 min and did not require expensive machine such as thermocycler, it can be used as a rapid and simple detection method for foodborne pathogens.

본 연구는 loop-mediated isothermal amplification(LAMP)을 이용하여 Clostridium difficile을 검출하고자 하였다. LAMP 수행을 위하여 선택적인 타깃 유전자로 C. difficile의 16S ribosomal RNA를 타깃으로 하여 primer set를 구성하였다. 5개의 primer set(BIP, FIP, B3, F3, LF, PF)를 이용하여 TcdA와 TcdB toxin이 모두 양성인 균주, TcdA와 TcdB toxin이 모두 음성인 균주와 식품 분리균주를 효과적으로 검출할 수 있었다. LAMP는 80분 이내의 시간이 필요하며 thermocycler와 같은 장비를 필요로 하지 않고 또한 결과를 직접 눈으로 확인할 수 있기 때문에 식품 생산 현장에서 활용될 수 있을 것이라고 생각되었다.

Keywords

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