Simultaneous Detection of Staphylococcus aureus, Salmonella enterica subsp., Vibrio parahaemolyticus by Multiplex Polymerase Chain Reaction

Multiplex Polymerase Chain Reaction(PCR)법을 이용한 Staphylococcus aureus, Salmonella enterica subsp., Vibrio parahaemolyticus의 다중동시검출

  • 정유석 ((재)대구테크노파크 바이오산업지원센터) ;
  • 정희경 ((재)대구테크노파크 바이오산업지원센터) ;
  • 전원배 (대구경북과학기술원) ;
  • 서화정 (대구경북과학기술원) ;
  • 홍주헌 (대구가톨릭대학교 외식식품산업학부)
  • Published : 2010.04.30


This study was conducted to detect and identify Staphylococcus aureus, Vibrio parahaemolyticus, and Salmonella enterica subsp. using simultaneous multiplex polymerase chain reaction (multiplex PCR) assay. 23S rRNA partial gene (S. aureus), tox R gene (V. parahaemolyticus), and inv A gene (S. enterica subsp.) as diagnostic marker gene were suggested, and their amplicon sizes were 482 bp, 368 bp, and 284 bp, respectively. Non specific amplicons by STA-5F/STA-5R primer, ToxR-F/ToxR-R primer, and 139/141 primer were not observed in genomic DNA of pathogen bacteria as Bacillus cereus, Listeria monocytogenes, Escherichia coli, Proteus vulgaris, Streptococcus pyogenes, Candida albicans, and Shigella sonnei. The extracted crude DNA of targeted bacteria was detected as PCR template successfully. The detection limits were $10^5\sim10^4$ CFU/mL and 10 pg of purified genomic DNA of S. aureus, V. parahaemolyticus, and S. enterica subsp. by using simultaneous multiplex PCR.


  1. Lee HJ. 2008. Pathogenic agents and outbreak of foodborne diseases at home and abroad. Kor J Vet Publ Hlth 32: 81-89.
  2. Jung HJ, Cho JI, Park SH, Ha SD, Lee KH, Kim CH, Song ES, Chung DH, Kim MG, Kim KY, Kim KS. 2005. Genotype and phenotype characteristics of Staphylococcus aureus isolates from lettuces and raw milk. Korean J Food SciTechnol 37: 134-141.
  3. Ingeborg H, Angelika L, Petra R, Kurt K, Ernst B, Marti W. 2001. Comparison of different approaches to quantify Staphylococcus aureus cells by real-time PCR and quantitative PCR and application of this technique for examination of cheese. Appl Environ Microbiol 67: 3122-3126.
  4. Le Loir Y, Baron F, Gautier M. 2003. Staphylococcus aureus and food poisoning. Genet Mol Res 2: 63-76.
  5. Yeh KS, Chen TH, Liao CW, Chang CS, Lo HC. 2002. PCR amplification of the Salmonella typhimurium fim Y gene sequence to detect the Salmonella species. Int J Food Microbiol 78: 227-234.
  6. Chiu TH, Chen TR, Hwang WZ, Tsen HY. 2005. Sequencing of an internal transcribed spacer region of 16S-23S rRNA gene and designing of PCR primers for the detection of Salmonella spp. in food. Int J Food Microbiol97: 259-265.
  7. Jung SJ, Kim HJ, Kim HY. 2005. Quantitative detection of Samonella typhimurium contamination in milk, using real time PCR. J Microbiol Biotechnol 15: 1353-1358.
  8. Park HJ, Kim HH, Park SH, Shin EG, Kim JH, Kim HY. 2008. Direct and quantitative analysis of Salmonella enterica serovar typhimurium using real time PCR from artificially contaminated chicken meat. J Microbiol Biotechnol18: 1453-1458.
  9. Oh EK, Yu HS, Shin SB, Son KR, Park KB, Kwon JY, Lee TS, Lee HJ. 2008. Trimethoprim resistance of Vibrio parahaemolyticus isolated from the fish farm. J Kor Fish Soc 41: 324-329.
  10. di Pinto A, Ciccarese G, de Corato R, Novello L, Terio V.2008. Detection of pathogenic Vibrio parahaemolyticus in southern Italian shelfish. Food Control 19: 1037-1041.
  11. Kim MM, Oh MH, Lee GY, Hwang IG, Kwak HS, Kang YS, Koh YH, Jun HK, Kwong KS. 2008. Analysis of major food borne pathogens in various foods in Korea. Food Sci Biotechnol 17: 483-488.
  12. Choi KY, Kim BS, Bae WS, Jung WS, Cho YJ. 2008.Developing the index of foodborne disease occurrence.Korean J Appl Statist 21: 649-658.
  13. Adndreja R, Benaissa EM, Mieke U, Philippe B, Willy Z,Ernst H, Ellen F, Johan D. 2006. Immunoquantitative real time PCR for detection and quantification of Staphylococcus aureus enterotoxin B in foods. Appl Environ Mircobiol 72: 16593-16599.
  14. Kwak HS, Lee DH, Moon HS, Park JS, Woo GJ, Kim CM. 2003. Evaluation of the efficiency of E. coli O157:H7 rapid detection kit using immunochromatography. J Fd Hyg Safety 18: 118-124.
  15. Kim MH, Kim WJ, Sin WS, Son DH, Cha SG. 2003. Feasibility study on the use of liposomes for detecting food-borne pathogenic bacteria. Korean J Food Sci AniResour 23: 278-283.
  16. Jung HJ, Cho JI, Song ES, Kim JJ, Kim KS. 2005. PCR detection of virulence genes encoding coagulase and other toxins among clinical methicillin-resistant Staphylococcus aureus isolates. Kor J Microbiol Biotechnol 33: 207-214.
  17. Lim JS, Yoon JH, Min BK, Hong KW. 2008. Detection and identification of shiga-like toxin producing Escherichia coli O157:H7 by multiplex PCR. Food Eng Prog 12: 8-14.
  18. Jeong SH, Kim MY, Kim HJ, Kim TU, Yu SL, Kim HY.2003. Rapid detection of Salmonella species in foods using PCR. J Korean Soc Agric Chem Biotechnol 46: 225-228.
  19. Han KH, Lee CW, Lee YS, Yang OS, Lim YK, Yoon BS. 2001. Application of multiplex PCR using Lis-mix primers in food test and specific detection of Listeria ivanovii . J Fd Hyg Safety 16: 251-257.
  20. Seo HA, Park SH, Kim GS. 2003. Optimization of the concentrations of ERIC-PCR components to simultaneously differentiate five foodborne pathogenic bacterial genera. J Fd Hyg Safety 18: 229-236.
  21. Kim SS, Lee HM, Lee JB. 2003. Molecular epidemiological characteristics of Vibrio parahaemolyticus as recently wilde-spreaded in Korea. Korean J Biotechnol Bioeng 18:522-528.
  22. Yoon JH, Lee SJ. Hong KW. 2008. A multiplex PCR Assay for the direct detection of Salmonella spp., Yersinia enterocolitica, and Staphylococcus aureus in Food. Food Eng Prog 12: 24-31.
  23. Park SH, Kim HJ, Kim HY. 2006. Simultaneous detection of Yersinia enterocolitica, Staphylococcus aureus, and Shigella spp. in lettuce using multiplex PCR method. J Microbiol Biotechnol 16: 1301-1305.

Cited by

  1. Development of a Novel PCR Primer for Detection of Staphylococcus aureus vol.49, pp.1, 2015,
  2. Rapid Detection for Salmonella spp. by Ultrafast Real-time PCR Assay vol.33, pp.1, 2018,