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http://dx.doi.org/10.9721/KJFST.2013.45.6.791

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

Kim, Hong-Seok (KU Center for Food Safety, College of Veterinary Medicine, Konkuk University)
Chon, Jung-Whan (KU Center for Food Safety, College of Veterinary Medicine, Konkuk University)
Kim, Dong-Hyeon (KU Center for Food Safety, College of Veterinary Medicine, Konkuk University)
Song, Kwang-Young (KU Center for Food Safety, College of Veterinary Medicine, Konkuk University)
Seo, Kun-Ho (KU Center for Food Safety, College of Veterinary Medicine, Konkuk University)
Publication Information
Korean Journal of Food Science and Technology / v.45, no.6, 2013 , pp. 791-795 More about this Journal
Abstract
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.
Keywords
Staphylococcus aureus; detection; culture method; PCR; livestock products;
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1 Pagadala S, Parveen S, Schwarz JG, Rippen T, Luchansky JB. Comparison of automated BAX PCR and standard culture methods for detection of Listeria monocytogenes in blue crabmeat (Callinectus sapidus) and blue crab processing plants. J. Food Protect. 74: 1930-1933 (2011)   DOI   ScienceOn
2 Shalaby MA, Mohamed MS, Mansour MA, Abd EHA. Comparison of polymerase chain reaction and conventional methods for diagnosis of Listeria monocytogenes isolated from different clinical specimens and food stuffs. Clin. Lab. 57: 919-924 (2011)
3 Malorny B, Huehn S, Dieckmann R, Krmer N, Helmuth R. Polymerase chain reaction for the rapid detection and serovar identification of Salmonella in food and feeding stuff. Food Anal. Method. 2: 81-95 (2009)   DOI
4 Hwang BH, Lee WJ, Cha HJ. Polymerase chain reaction-based detection of total and specific Vibrio species. Appl. Biochem. Biotech. 162: 1187-1194 (2010)   DOI   ScienceOn
5 Chon JW, Park JS, Hyeon JY, Park C, Song KY, Hong KW, Hwang IG, Kwak HS, Seo KH. Development of real-time PCR for the detection of Clostridium perfringens in meats and vegetables. J. Microbiol. Biotechn. 22: 530-534 (2012)   과학기술학회마을   DOI   ScienceOn
6 Rushdy AA, Salama MS, Othman AS. Detection of methicillin/ oxacillin resistant Staphylococcus aureus isolated from some clinical hospitals in Cairo using Meca/Nuc genes and antibiotic susceptibility profile. Int. J. Agric. Biol. 9: 800-806 (2007)
7 Thomas LC, Gidding HF, Ginn AN, Olma T, Iredell J. Development of a real-time Staphylococcus aureus and MRSA (SAM-) PCR for routine blood culture. J. Microbiol. Meth. 68: 296-302 (2007)   DOI   ScienceOn
8 Riyaz-Ul-Hassan S, Verma V, Qazi GN. Evaluation of three different molecular markers for the detection of Staphylococcus aureus by polymerase chain reaction. Food Microbiol. 25: 452-459 (2008)   DOI   ScienceOn
9 Straub JA, Hertel C, Hammes WP. A 23S rRNA-targeted polymerase chain reaction-based system for detection of Staphylococcus aureus in meat starter cultures and dairy products. J. Food Protect. 62: 1150-1156 (1999)
10 Maes N, Magdalena J, Rottiers S, De Gheldre Y, Struelens MJ. Evaluation of a triplex PCR assay to discriminate Staphylococcus aureus from coagulase-negative staphylococci and determine methicillin resistance from blood cultures. J. Clin. Microbiol. 40: 1514-1517 (2002)   DOI   ScienceOn
11 Chikkala R, George NO, Ratnakar KS, Iyer RN, Sritharan V. Heterogeneity in femA in the indian isolates of Staphylococcus aureus limits its usefulness as a species specific marker. Adv. Infect. Dis. 2: 82-88 (2012)   DOI
12 Jonas D, Grundmann H, Hartung D, Daschner FD, Towner KJ. Evaluation of the mecA femB duplex polymerase chain reaction for detection of methicillin-resistant Staphylococcus aureus. Eur. J. Clin. Microbiol. 18: 643-647 (1999)   DOI   ScienceOn
13 Stephan R, Annemller C, Hassan AA, Lmmler C. Characterization of enterotoxigenic Staphylococcus aureus strains isolated from bovine mastitis in north-east Switzerland. Vet. Microbiol. 78: 373-382 (2001)   DOI   ScienceOn
14 Pinto B, Chenoll E, Aznar R. Identification and typing of food-borne Staphylococcus aureus by PCR-based techniques. Syst. Appl. Microbiol. 28: 340-352 (2005)   DOI   ScienceOn
15 Seo KH, Brackett RE. Rapid, specific detection of Enterobacter sakazakii in infant formula using a real-time PCR assay. J. Food Protect. 68: 59-63 (2005)
16 Chon JW, Song KY, Kim SY, Hyeon JY, Kim YG, Hwang IG, Kwak HS, Seo KH. Comparison of real-time PCR and conventional culture method for detection of Cronobacter spp. in powdered foods. Korean J. Microbiol. 47: 87-91 (2011)   과학기술학회마을
17 Lee JH, Song KY, Hyeon JY, Hwang IG, Kwak HS, Han JA, Chung YH, Seo KH. Comparison of standard culture method and real-time PCR assay for detection of Staphylococcus aureus in processed and unprocessed foods. Korean J. Food Sci. Ani. Resour. 30: 410-418 (2010)   과학기술학회마을   DOI   ScienceOn
18 Hyeon JY, Hwang IG, Kwak HS, Park JS, Heo S, Choi IS, Park C, Seo KH. 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. Korean J. Food Sci. Ani. Resour. 29: 506-512 (2009)   과학기술학회마을   DOI   ScienceOn
19 Han SR, Hyeon JY, Kim HY, Park JS, Heo S, Shin HC, Seo KH. Evaluation of conventional culture methods and validation of immunoassays for rapid detection of Listeria monocytogenes in dairy and processed foods. Korean J. Food Sci. Ani. Resour. 28: 616-622 (2008)   과학기술학회마을   DOI   ScienceOn
20 Brakstad OG, Aasbakk K, Maeland JA. Detection of Staphylococcus aureus by polymerase chain reaction amplification of the nuc gene. J. Clin. Microbiol. 30: 1654-1660 (1992)
21 De Buyser ML, Lombard B, Schulten SM, In't Veld PH, Scotter SL, Rollier P, Lahellec C. Validation of EN ISO standard methods 6888 Part 1 and Part 2: 1999-Enumeration of coagulase-positive staphylococci in foods. Int. J. Food Microbiol. 83: 185-194 (2003)   DOI   ScienceOn
22 Kim MH, Kim WJ, Shin WS, Shon DH, Cha SK. Feasibility study on the use of liposomes for detecting food-borne pathogenic bacteria. Korean J. Food Sci. Ani. Resour. 23: 278-283 (2003)   과학기술학회마을
23 Davis JA, Farrah SR, Wilkie AC. Selective growth of Staphylococcus aureus from flushed dairy manure wastewater using acriflavinesupplemented mannitol salt agar. Lett. Appl. Microbiol. 42: 606-611 (2006)
24 Bickley J, Short JK, McDowell DG, Parkes HC. Polymerase chain reaction (PCR) detection of Listeria monocytogenes in diluted milk and reversal of PCR inhibition caused by calcium ions. Lett. Appl. Microbiol. 22: 153-158 (1996)   DOI   ScienceOn
25 Opel KL, Chung D, McCord BR. A study of PCR inhibition mechanisms using real time PCR. J. Forensic Sci. 55: 25-33 (2010)   DOI   ScienceOn
26 Warren BR, Parish ME, Schneider KR. Comparison of conventional culture methods and fta filtration-nested PCR for the detection of Shigella boydii and Shigella sonnei on tomato surfaces. J. Food Protect. 68: 1606-1612 (2005)
27 Alarcon B, Vicedo B, Aznar R. PCRbased procedures for detection and quantification of Staphylococcus aureus and their application in food. J. Appl. Microbiol. 100: 352-364 (2006)   DOI   ScienceOn
28 Bae HJ, Park HJ. Hazard analysis of Staphylococcus aureus in ready-to-eat sandwiches. J. Korean Soc. Food Sci. Nutr. 36: 938-943 (2007)   과학기술학회마을   DOI   ScienceOn
29 QIA. The Processing Standards and Ingredient Specifications for Livestock Products. Korean Animal and Plant Quarantine Agency. Anyang, Korea. pp. 203-205 (2012)