Journal of Food Hygiene and Safety (한국식품위생안전성학회지)

The Korean Society of Food Hygiene and Safety (한국식품위생안전성학회)
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Comparison of Loop-Mediated Isothermal Amplification and Real-Time PCR for the Rapid Detection of Salmonella Typhimurium, Listeria monocytogenes and Cronobacter sakazakii Artificially Inoculated in Foods

식품에 인위접종된 Salmonella Typhimurium, Listeria monocytogenes, Cronobacter sakazakii의 신속검출을 위한 Real-time PCR과 Loop-mediated isothermal amplification 비교

  • Kim, Jin-Hee (Department of Food and Nutrition, Kookmin University) ;
  • Oh, Se-Wook (Department of Food and Nutrition, Kookmin University)
  • 김진희 (국민대학교 식품영양학과) ;
  • 오세욱 (국민대학교 식품영양학과)
  • Received : 2018.12.07
  • Accepted : 2019.03.13
  • Published : 2019.04.30
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Abstract

The objective of this research was to compare loop-mediated isothermal amplification (LAMP) with real-time polymerase chain reaction (PCR) for the rapid detection of pathogens in foods. In this study, the limits of detection (LODs) for Salmonella Typhimurium, Listeria monocytogenes, and Cronobacter sakazakii were evaluated in various foods. Among 11 samples tested for S. Typhimurium, LAMP and real-time PCR had the same LODs in beef and duck meat whereas real-time PCR was more sensitive than the LAMP in 8 samples. However, S. Typhimurium in chocolate samples was not detected by real-time PCR. The sensitivity of real-time PCR was high in all samples inoculated with L. monocytogenes and C. sakazakii whereas LAMP was more sensitive than real-time PCR in oil-rich foods. Therefore, LAMP can be shown as an easrer, more convenient method, as well as effective analytical method for testing difficult samples when employed in PCR.

식품에 존재하는 병원균을 신속검출하기 위한 방법으로 LAMP와 real-time PCR 방법을 비교 평가 하였다. S. Typhimurium, L. monocytogenes, C. sakazakii의 3종에 대해 식품공전에서 권고하는 식품 종류를 선별하여 민감도를 분석하였다. S. Typhimurium에서는 11종의 식품(햄, 닭가슴살, 계란, 돼지고기, 소고기, 오리고기, 액상음료, 샐러드, 콘프레이크, 초콜릿, 사료)중 4종(햄, 돼지고기, 시리얼, 사료)에서 LAMP보다 real-time PCR에서 검출 민감도가 10배 이상 더 높았고, 6종(닭가슴살, 계란, 소고기, 오리고기, 음료, 샐러드)에서는 real-time PCR과 비슷한 수준을 그리고 초콜릿에서는 real-time PCR로는 검출되지 않았으며 LAMP로만 검출되는 결과가 나타났다. L. monocytogenes와 C. sakazakii에서는 9종 모두에서 LAMP보다 real-time PCR에서 검출 민감도가 더 높았다. 또한 L. monocytogenes에서 LAMP의 검출 민감도가 S. Typhimurium과 C. sakazakii 보다 10배 이상 낮았다. 3M MDS의 검출한계 향상을 위해 변형된 3M MDS의 민감도는 기존대비 10배 이상 증가되었다. 따라서 식품에 존재하는 병원균의 검출을 위해 식품의 구성성분에 따라 LAMP와 real-time PCR를 적절히 선택하는 것이 바람직할 것으로 생각되었다. 한편, 농축 방법을 이용해 LAMP방법의 민감도를 향상시킬 수 있음을 알 수 있었다.

Keywords

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