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Comparison of Standard Culture Method and Real-time PCR Assay for Detection of Staphylococcus aureus in Processed and Unprocessed Foods

가공식품과 비가공식품에서의 황색포도상구균 검출을 위한 배지법과 Real-time PCR법의 비교

  • Lee, Jae-Hoon (Department of Public Health, College of Veterinary Medicine, Konkuk University) ;
  • Song, Kwang-Young (Department of Public Health, College of Veterinary Medicine, Konkuk University) ;
  • Hyeon, Ji-Yeon (Department of Public Health, College of Veterinary Medicine, Konkuk University) ;
  • Hwang, In-Gyun (Korea Food and Drug Administration, National Institute of Food and Drug Safety Evaluation) ;
  • Kwak, Hyo-Sun (Korea Food and Drug Administration, National Institute of Food and Drug Safety Evaluation) ;
  • Han, Jeong-A (Korea Food and Drug Administration, National Institute of Food and Drug Safety Evaluation) ;
  • Chung, Yun-Hee (Korea Consumer Agency) ;
  • Seo, Kun-Ho (Department of Public Health, College of Veterinary Medicine, Konkuk University)
  • 이재훈 (건국대학교 수의과대학) ;
  • 송광영 (건국대학교 수의과대학) ;
  • 현지연 (건국대학교 수의과대학) ;
  • 황인균 (식품의약품안전청 식품의약품안전평가원) ;
  • 곽효선 (식품의약품안전청 식품의약품안전평가원) ;
  • 한정아 (식품의약품안전청 식품의약품안전평가원) ;
  • 정윤희 (한국소비자원) ;
  • 서건호 (건국대학교 수의과대학)
  • Received : 2009.11.23
  • Accepted : 2010.03.19
  • Published : 2010.06.30

Abstract

Staphylococcus aureus is one of the major pathogens that can cause staphylococcal infection and food poisoning. In this study, we compared conventional culture methods and real-time PCR for detection of S. aureus in artificially inoculated milk, sausage, raw pork, and vegetable salad. The performance of a coagulase test for confirming S. aureus was also compared with a colony PCR test. Bulk food samples (500 g each) were artificially inoculated with S. aureus and divided into 20 samples (25 g or mL each). All samples were added to tryptic soy broth (225 mL/sample) with 10% NaCl and incubated at $37^{\circ}C$ for 24 h. After the enrichment, broth cultures were streaked onto Baird-Parker (BP) agar with egg yolk tellulite, and incubated at $37^{\circ}C$ for 24 h. In addition, 1 mL of broth cultures was collected to perform real-time PCR. Two suspicious colonies from the BP agar were picked up and plated on nutrient agar and incubated at $37^{\circ}C$ for 24 h followed, by a coagulase confirmation test and a colony PCR analysis. There were no statistical differences between culture methods and realtime PCR in food samples with low background microflora, such as milk and sausage. However, a significant statistical difference was found between the culture methods and real-time PCR for raw pork and vegetable salad. Furthermore, the colony PCR test of the presumptive colonies on BP agar for confirming S. aureus is more accurate and efficient than the coagulase test for unprocessed foods.

Keywords

standard culture methods;real-time PCR;foods

Acknowledgement

Supported by : 건국대학교

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