Food Science of Animal Resources (한국축산식품학회지)

Korean Society for Food Science of Animal Resources (한국축산식품학회)
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Evaluation of an Automated ELISA (VIDAS(R)) and Real-time PCR by Comparing with a Conventional Culture Method for the Detection of Salmonella spp. in Steamed Pork and Raw Broccoli Sprouts

편육과 브로콜리싹에서 Salmonella spp. 검출을 위한 배지법과 Real-time PCR 및 신속 검사키트(VIDAS(R))의 비교검증

  • Hyeon, Ji-Yeon (Department of Public Health, College of Veterinary Medicine, Konkuk University) ;
  • Hwang, In-Gyun (Food Microbiology Division, Food Safety Evaluation Department, National Institute of Food and Drug Safety Evaluation) ;
  • Kwak, Hyo-Sun (Food Microbiology Division, Food Safety Evaluation Department, National Institute of Food and Drug Safety Evaluation) ;
  • Park, Jong-Seok (Research Planning & Management Division, National Institute of Food and Drug Safety Evaluation) ;
  • Heo, Seok (Research Planning & Management Division, National Institute of Food and Drug Safety Evaluation) ;
  • Choi, In-Soo (Department of Infectious Diseases, College of Veterinary Medicine, KonKuk University) ;
  • Park, Chan-Kyu (Department of Animal Biotechnology, College of Animal Bioscience and Technology, Konkuk University) ;
  • Seo, Kun-Ho (Department of Public Health, College of Veterinary Medicine, Konkuk University)
  • 현지연 (건국대학교 수의과대학 공중보건학) ;
  • 황인균 (식품의약품안전청 식품의약품안전평가원 미생물과) ;
  • 곽효선 (식품의약품안전청 식품의약품안전평가원 미생물과) ;
  • 박종석 (식품의약품안전청 식품의약품안전평가원 연구기획조정과) ;
  • 허석 (식품의약품안전청 식품의약품안전평가원 연구기획조정과) ;
  • 최인수 (건국대학교 수의과대학 전염병학) ;
  • 박찬규 (건국대학교 동물생명과학대학) ;
  • 서건호 (건국대학교 수의과대학 공중보건학)
  • Published : 2009.08.31
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Abstract

Salmonellosis is an important worldwide foodborne infectious disease that is transmitted by many food vehicles including raw and processed animal products and fresh produce. In this study, the effectiveness of automated ELISA ($VIDAS^{(R)}$) and realtime PCR in the detection of Salmonella spp. in steamed pork and raw broccoli sprouts was evaluated by comparing their results with those of a conventional culture method. Bulk samples (500 g) of steamed pork and raw broccoli sprouts were inoculated with various levels of Salmonella and divided into 20 samples (25 g each). All the samples, including the controls, were analyzed using a conventional culture method, $VIDAS^{(R)}$, and real-time PCR to detect the presence of Salmonella. In addition, the levels of background flora in the steamed pork and the raw broccoli sprouts were determined. In the steamed pork that contained less than 100 CFU/g of aerobic bacteria, all three methods detected low levels of Salmonella without a statistical difference in their performance. In the broccoli sprouts with high quantities of background flora (ca. $6.7{\times}10^7$ CFU/g), however, all three methods were unable to detect low levels of Salmonella, and real-time PCR and $VIDAS^{(R)}$ more sensitively detected Salmonella than the culture method, with significant statistical differences. In conclusion, $VIDAS^{(R)}$ and real-time PCR could be superior to conventional culture methods in detecting Salmonella in food with high levels of background flora.

본 연구에서는 편육과 브로콜리 싹에 Salmonella 농도를 단계별로 접종하고 배지법, $VIDAS^{(R)}$, real-time PCR의 양성 검출률을 비교함으로써 $VIDAS^{(R)}$, real-time PCR의 Salmonella 검출력을 검증하였으며, 또한 각 식품별로 검출률에 차이가 있는가를 알아보았다. 편육과 브로콜리 싹(500 g)에 3단계 농도(<15, 15-100, 100-1000 CFU/500 g)를 접종하고 20개 샘플로 나눈 후 배지법, $VIDAS^{(R)}$ Salmonella ($VIDAS^{(R)}$ SLM), real-time PCR법을 시행하여 검출능력을 비교하였다. 편육에서 배지법, $VIDAS^{(R)}$, real-time PCR를 이용하여 Salmonella를 검출한 결과, 낮은 접종 농도(<15 CFU/500 g)에서도 Salmonella가 검출되었고 $VIDAS^{(가)}$O real-time PCR 모두 배지법과 통계학적 유의차가 나타나지 않았다. 반면에 브로콜리 싹을 이용하였을 때 낮은 접종 농도에서는 Salmonella가 검출되지 않았으며 $VIDAS^{(가)}$O real-time PCR 모두 배지법보다 더 많은 샘플을 검출하였다. 따라서 정상 세균총이 많은 야채 식품의 경우 배지법은 경쟁 세균에 의해 검출이 저해되므로 이보다 영향을 덜 받는 $VIDAS^{(R)}$법이나 real-time PCR법을 이용하여 검출하는 것이 더 효과적이라고 할 수 있겠다. 또한 이러한 신속 진단법을 이용하면 $VIDAS^{(R)}$는 3일, real-time PCR은 1일 이내에 배지법과 같거나 보다 높은 검출률로 Salmonella를 검출할 수 있는 것으로 나타났다.

Keywords

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