2013•Korean Journal of Food Science and TechnologyOpen access

Evaluation of a PCR Assay for the Rapid Detection of Staphylococcus aureus in Milk and Meat Products

Hong-Seok Kim, Jung‐Whan Chon, Dong‐Hyeon Kim, Kwang‐Young Song, Kun‐Ho Seo

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Abstract

본 연구에서는 유제품 및 육제품에서 Staphylococcus aureus를 검출하기 위한 PCR과 배지배양법을 비교검증 하였다. 우유, 분유, 소시지, 소고기 분쇄육에 S. aureus를 인위적으로 접종시킨 후, 축산물 가공기준 및 성분규격에 따라 10% NaCl이 첨가된 tryptic soy broth(TSB)에서 증균배양 하였으며, Baird Parker agar에 선택배양 하였다. PCR은 TSB에서 1 mL를 채취한 후 DNA를 추출하여 시행하였으며, S. aureus의 23s rRNA를 표적으로 하는 primer를 사용하였다. 의심집락에 대해 coagulase test와 colony PCR을 통한 확인시험을 실시하였다. 실험결과 우유와 분유에서는 배지배양법과 PCR간에 통계학적 유의차가 존재하지 않았다. 소시지와 소고기 분쇄육의 경우에는 PCR이 배지법에 비해 더 많은 양성을 검출하여 높은 통계학적 유의차를 보였다(p<0.05). 연구결과를 종합하면, 유제품 및 육제품에서 PCR을 통한 S. aureus의 검출은 기존 배지배지배양법에 비해 시간과 노동력을 절감할 수 있는 효과적인 방법으로 나타났다. The aim of this study was to compare the performance of a standard culture method and polymerase chain reaction (PCR) for the detection of Staphylococcus aureus (S. aureus) in milk and meat products. Milk, dried infant formula, sausage and ground beef that had been artificially inoculated with S. aureus were enriched in tryptic soy broth. After the enrichment, a loopful was inoculated onto Baird-Parker agar with egg-yolk-tellurite. In parallel, 23S rRNA was amplified by PCR from samples of the enriched broth. Suspected S. aureus colonies grown on selective agars were finally confirmed by a coagulase test and colony PCR. No significant statistical differences were observed between the incidence of S. aureus detected by the culture method and the incidence detected by PCR, in milk or dried infant formula. However, in sausage and ground beef, the number of positives detected by PCR was significantly higher than by the culture method (p<0.05). Our findings suggest that PCR could be an effective screening tool for the detection of S. aureus compared to the standard culture method.

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본 연구에서는 유제품 및 육제품에서 Staphylococcus aureus를 검출하기 위한 PCR과 배지배양법을 비교검증 하였다. 우유, 분유, 소시지, 소고기 분쇄육에 S. aureus를 인위적으로 접종시킨 후, 축산물 가공기준 및 성분규격에 따라 10% NaCl이 첨가된 tryptic soy broth(TSB)에서 증균배양 하였으며, Baird Parker agar에 선택배양 하였다. PCR은 TSB에서 1 mL를 채취한 후 DNA를 추출하여 시행하였으며, S. aureus의 23s rRNA를 표적으로 하는 primer를 사용하였다. 의심집락에 대해 coagulase test와 colony PCR을 통한 확인시험을 실시하였다. 실험결과 우유와 분유에서는 배지배양법과 PCR간에 통계학적 유의차가 존재하지 않았다. 소시지와 소고기 분쇄육의 경우에는 PCR이 배지법에 비해 더 많은 양성을 검출하여 높은 통계학적 유의차를 보였다(p<0.05). 연구결과를 종합하면, 유제품 및 육제품에서 PCR을 통한 S. aureus의 검출은 기존 배지배지배양법에 비해 시간과 노동력을 절감할 수 있는 효과적인 방법으로 나타났다. The aim of this study was to compare the performance of a standard culture method and polymerase chain reaction (PCR) for the detection of Staphylococcus aureus (S. aureus) in milk and meat products. Milk, dried infant formula, sausage and ground beef that had been artificially inoculated with S. aureus were enriched in tryptic soy broth. After the enrichment, a loopful was inoculated onto Baird-Parker agar with egg-yolk-tellurite. In parallel, 23S rRNA was amplified by PCR from samples of the enriched broth. Suspected S. aureus colonies grown on selective agars were finally confirmed by a coagulase test and colony PCR. No significant statistical differences were observed between the incidence of S. aureus detected by the culture method and the incidence detected by PCR, in milk or dried infant formula. However, in sausage and ground beef, the number of positives detected by PCR was significantly higher than by the culture method (p<0.05). Our findings suggest that PCR could be an effective screening tool for the detection of S. aureus compared to the standard culture method.

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Available abstract

본 연구에서는 유제품 및 육제품에서 Staphylococcus aureus를 검출하기 위한 PCR과 배지배양법을 비교검증 하였다. 우유, 분유, 소시지, 소고기 분쇄육에 S. aureus를 인위적으로 접종시킨 후, 축산물 가공기준 및 성분규격에 따라 10% NaCl이 첨가된 tryptic soy broth(TSB)에서 증균배양 하였으며, Baird Parker agar에 선택배양 하였다. PCR은 TSB에서 1 mL를 채취한 후 DNA를 추출하여 시행하였으며, S. aureus의 23s rRNA를 표적으로 하는 primer를 사용하였다. 의심집락에 대해 coagulase test와 colony PCR을 통한 확인시험을 실시하였다. 실험결과 우유와 분유에서는 배지배양법과 PCR간에 통계학적 유의차가 존재하지 않았다. 소시지와 소고기 분쇄육의 경우에는 PCR이 배지법에 비해 더 많은 양성을 검출하여 높은 통계학적 유의차를 보였다(p<0.05). 연구결과를 종합하면, 유제품 및 육제품에서 PCR을 통한 S. aureus의 검출은 기존 배지배지배양법에 비해 시간과 노동력을 절감할 수 있는 효과적인 방법으로 나타났다. The aim of this study was to compare the performance of a standard culture method and polymerase chain reaction (PCR) for the detection of Staphylococcus aureus (S. aureus) in milk and meat products. Milk, dried infant formula, sausage and ground beef that had been artificially inoculated with S. aureus were enriched in tryptic soy broth. After the enrichment, a loopful was inoculated onto Baird-Parker agar with egg-yolk-tellurite. In parallel, 23S rRNA was amplified by PCR from samples of the enriched broth. Suspected S. aureus colonies grown on selective agars were finally confirmed by a coagulase test and colony PCR. No significant statistical differences were observed between the incidence of S. aureus detected by the culture method and the incidence detected by PCR, in milk or dried infant formula. However, in sausage and ground beef, the number of positives detected by PCR was significantly higher than by the culture method (p<0.05). Our findings suggest that PCR could be an effective screening tool for the detection of S. aureus compared to the standard culture method.

Key concepts: Staphylococcus aureus, Tryptic soy broth, Microbiology, Agar, Coagulase, Biology, Polymerase chain reaction, Food science

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