Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Expression of Enterotoxin Genes in Staphylococcus aureus Isolates Based on mRNA Analysis

  • Lee, Young-Duck (School of Life Science and Biotechnology, Korea University) ;
  • Moon, Bo-Youn (Food Safety Research Division, Korea Food Research Institute) ;
  • Park, Jong-Hyun (Department of Food Science and Biotechnology, Kyungwon University) ;
  • Chang, Hyo-Ihl (School of Life Science and Biotechnology, Korea University) ;
  • Kim, Wang-June (Department of Food Science and Technology, Dongguk University)
  • Published : 2007.03.31
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Abstract

Staphylococcus aureus strains are important foodborne pathogens that produce various toxins. To evaluate the risk of the enterotoxins, four S. aureus strains from kimbap and two clinical samples were isolated and identified, and their expression of the enterotoxin genes were analyzed using a reverse transcription real-time PCR. Various enterotoxin genes were detected, including sea, seg, seh, sei, sen, seo, and sem, where each isolate contained one or two. When the mRNA detection of the enterotoxin genes was analyzed using a reverse transcriptase PCR, various levels of expression were found depending on the species and enterotoxin gene. Therefore, it is reasonable to suggest that the poisoning risk of S. aureus can be effectively evaluated based on the gene expression at the mRNA level.

Keywords

References

  1. Altboum, Z., I. Hertman, and S. Sarid. 1990. Penicillinase plasmid-linked genetic determinants for enterotoxins B and C1 production in Staphylococcus aureus. Mol. Gen. Genet. 220: 329-333
  2. Altekruse, S. F., M. L. Cohen, and D. L. Swerdlow. 1997. Emerging foodborne diseases. Emerg. Infect. Dis. 3: 285- 293 https://doi.org/10.3201/eid0303.970304
  3. Balaban, N. and A. Rasooly. 2000. Staphylococcal enterotoxins. Int. J. Food Microbiol. 61: 1-10 https://doi.org/10.1016/S0168-1605(00)00377-9
  4. Bayles, K. W. and J. J. Iandolo. 1989. Genetic and molecular analyses of the gene encoding staphylococcal enterotoxin D. J. Bacteriol. 171: 4799-4806 https://doi.org/10.1128/jb.171.9.4799-4806.1989
  5. Bergdoll, M. S. 1989. Staphylococcus aureus. In: Foodborne Bacterial Pathogens, pp. 463-523. Marcel Dekker, Inc., New York, U.S.A
  6. Betley, M. J. and J. J. Mekalanos. 1985. Staphylococcal enterotoxin A is encoded by phage. Science 229: 185-187 https://doi.org/10.1126/science.3160112
  7. Blaiotta, G., D. Ercolini, C. Pennacchia, V. Fusco, A. Casaburi, O. Pepe, and F. Villani. 2004. PCR detection of staphylococcal enterotoxin genes in Staphylococcus spp. strains isolated from meat and dairy products. J. Appl. Microbiol. 97: 719-730 https://doi.org/10.1111/j.1365-2672.2004.02349.x
  8. Boerema, J. A., R. Clemens, and G. Brightwell. 2006. Evaluation of molecular methods to determine enterotoxigenic status and molecular genotype of bovine, ovine, human and food isolates of Staphylococcus aureus. Int. J. Food Microbiol. 107: 192-201 https://doi.org/10.1016/j.ijfoodmicro.2005.07.008
  9. Bohach, G. A. and P. M. Schlievert. 1989. Conservation of the biologically active portions of staphylococcal enterotoxins C1 and C2. Infect. Immun. 57: 2249-2252
  10. Borst, D. W. and M. J. Betley. 1994. Promoter analysis of the staphylococcal enterotoxin A gene. J. Biol. Chem. 269: 1883-1888
  11. Bunning, V. K., J. A. Lindsay, and D. L. Archer. 1997. Chronic health effects of microbial foodborne disease. World Health Stat. 50: 51-56
  12. Chang, H. C. and M. S. Bergdoll. 1979. Purification and some physicochemical properties of staphylococcal enterotoxin D. Biochemistry 18: 1937-1942 https://doi.org/10.1021/bi00577a013
  13. Chen, T. R., C. S. Chiou, and H. Y. Tsen. 2004. Use of novel PCR primers specific to the genes of staphylococcal enterotoxin G, H, I for the survey of Staphylococcus aureus strains isolated from food-poisoning cases and food samples in Taiwan. Int. J. Food Microbiol. 92: 189-197 https://doi.org/10.1016/j.ijfoodmicro.2003.10.002
  14. Cheung, A. L., K. J. Eberhardt, and V. A. Fischetti. 1994. A method to isolate RNA from Gram-positive bacteria and mycobacteria. Anal. Biochem. 222: 511-514 https://doi.org/10.1006/abio.1994.1528
  15. Christiane, G., G. B. Manfred, and W. Christiane. 2001. Quantification of bacterial transcripts during infection using competitive reverse transcription-PCR (RT-PCR) and LightCycler RT-PCR. Clin. Diagn. LAB. Immunol. 8: 279- 282
  16. Couch, J. L., M. T. Soltis, and M. J. Betley. 1988. Cloning and nucleotide sequence of the type E staphylococcal enterotoxin gene. J. Bacteriol. 170: 2954-2960 https://doi.org/10.1128/jb.170.7.2954-2960.1988
  17. Cremonesi, P., M. Luzzana, M. Brasca, S. Morandi, R. Lodi, C. Vimercati, D. Agnellini, G. Caramenti, P. Moroni, and B. Castiglioni. 2005. Development of a multiplex PCR assay for the identification of Staphylococcus aureus enterotoxigenic strains isolated from milk and dairy products. Mol. Cell. Probes 19: 299-305 https://doi.org/10.1016/j.mcp.2005.03.002
  18. Holmberg, S. D. and P. A. Blake. 1984. Staphylococcal food poisoning in the United States. New facts and old misconceptions. JAMA 251: 487-489 https://doi.org/10.1001/jama.251.4.487
  19. Hovde, C. J., S. P. Hackett, and G. A. Bohach. 1990. Nucleotide sequence of the staphylococcal enterotoxin C3 gene: Sequence comparison of all three type C staphylococcal enterotoxins. Mol. Gen. Genet. 220: 329- 333
  20. Huang, I. Y., J. L. Hughes, M. S. Bergdoll, and E. J. Schantz. 1987. Complete amino acid sequence of staphylococcal enterotoxin A. J. Biol. Chem. 262: 7006-7013
  21. Jarraud, S., C. Mougel, J. Thiolouse, G. Lina, H. Meugnier, F. Forey, X. Nesme, J. Etienne, and F. Vandenesch. 2002. Relationships between Staphylococcus aureus genetic background, virulence factors, agr groups(alleles), and human disease. Infect. Immun. 70: 631-641 https://doi.org/10.1128/IAI.70.2.631-641.2002
  22. Johnson, W., M. S. Tyler, S. D. Ewan, E. P. Ashton, F. E. Polland, and K. R. Rozee. 1991. Detection of genes for enterotoxins, exofoliative toxins and toxic shock syndrome toxin 1 in Staphylococcus aureus by polymerase chain reaction. J. Clin. Microbiol. 29: 426-430
  23. Jung, H. J., K. S. Choi, and D. G. Lee. 2005. Synergistic killing effect of synthetic peptide P20 and cefotaxime on methicillin-resistant nocosomical isolate of Staphylococcus aureus. J. Microbiol. Biotechnol. 15: 1039-1046
  24. Kim, T. 2001. The economic impact of methicillin-resistant Staphylococcus aureus in Canadian hospitals. Infect. Cont. Hosp. Epidemiol. 22: 99-104 https://doi.org/10.1086/501871
  25. KFDA. Food Code. 2002. Korea Food and Drug Administration, Seoul, Korea
  26. Lee, D. H., B. J. Park, M. S. Lee, J. B. Choi, J. K. Kim, J. H. Park, and J. C. Park. 2006. Synergistic effect of Staphylococcus aureus and LPS on silica-induced tumor necrosis factor production in macrophage cell line J774A. J. Microbiol. Biotechnol. 16: 136-140
  27. Levine, W. C., J. F. Smart, D. L. Archer, N. H. Bean, and R. V. Tauxe. 1991. Foodborne disease outbreaks in nursing homes, 1975 through 1987. JAMA 16: 2105-2109
  28. Le Loir, Y., F. Baron, and M. Gautier. 2003. Staphylococcus aureus and food poisoning. Genet. Mol. Res. 31: 63-76
  29. Monday, S. R. and G. A. Bohach. 1999. Use of multiplex PCR to detect classical and newly described pyrogenic toxin genes in staphylococcal isolates. J. Clin. Microbiol. 37: 3411-3414
  30. McLauchlin, J., G. L. Narayanan, V. Mithani, and G. O'Neil. 2000. The detection of enterotoxins and toxic shock syndrome genes in Staphylococcus aureus by polymerase chain reaction. J. Food Prot. 63: 479-488 https://doi.org/10.4315/0362-028X-63.4.479
  31. Munson, S. H., M. T. Tremaine, M. J. Betley, and R. A. Welch. 1998. Identification and characterization of staphylococcal enterotoxin types G and I from Staphylococcus aureus. Infect. Immun. 66: 3337-3348
  32. Najera-Sanchez, G., R. Maldonado-Rodriguez, P. Ruiz Olvera, and L. M. de la Garza. 2003. Development of two multiplex polymerase chain reactions for the detection of enterotoxigenic strains of Staphylococcus aureus isolated from foods. J. Food Prot. 66: 1055-1062 https://doi.org/10.4315/0362-028X-66.6.1055
  33. Orwin, P. M., D. Y. Leung, T. J. Tripp, G. A. Bohach, C. A. Earhart, D. H. Ohlendorf, and P. M. Schlievert. 2002. Characterization of a novel staphylococcal enterotoxinlike superantigen, a member of the group V subfamily of pyrogenic toxins. Biochemistry 41: 14033-14040 https://doi.org/10.1021/bi025977q
  34. Orlando, C., P. Pinzani, and M. Pazzagli. 1998. Developments in quantitative PCR. Clin. Chem. Lab. Med. 36: 255-269 https://doi.org/10.1515/CCLM.1998.045
  35. Park, C. E., D. Warburton, and P. J. Laffey. 1996. A collaborative study on the detection of staphylococcal enterotoxins in foods with an enzyme immunoassay kit (TECRA). J. Food Prot. 59: 390-397 https://doi.org/10.4315/0362-028X-59.4.390
  36. Park, S. H., H. J. Kim, and H. Y. kim. 2006. Simultaneous detection of Yersinia enterocolitica, Staphylococcus aureus and Shigella spp. in lettuce using multiplex PCR method. J. Microbiol. Biotechnol. 16: 1301-1305
  37. Ramesh, A., B. P. Padmapriya, A. Chrashekar, and M. C. Varadaraj. 2002. Application of a convenient DNA extraction method and multiplex PCR for the direct detection of Staphylococcus aureus and Yersinia enterocolitica in milk samples. Mol. Cell. Probes 16: 307-314 https://doi.org/10.1006/mcpr.2002.0428
  38. Ren, K., J. D. Bannan, V. Pancholi, A. L. Cheung, J. C. Robbins, V. A. Fischetti, and J. B. Zabriskie. 1994. Characterization and biological properties of a new staphylococcal exotoxin. J. Exp. Med. 180: 1675-1683 https://doi.org/10.1084/jem.180.5.1675
  39. Rosec, J. P. and O. Gigaud. 2002. Staphylococcal enterotoxin genes of classical and new types detected by PCR in France. Int. J. Food Microbiol. 77: 61-70 https://doi.org/10.1016/S0168-1605(02)00044-2
  40. Saleema, S. L., M. Michelle, G. K. S. Rachel, C. E. Parastu, M. M. Hart, and J. T. Trevors. 2005. Microbial gene expression in soil: Methods, applications and challenges. J. Microbiol. Method. 63: 1-19 https://doi.org/10.1016/j.mimet.2005.03.007
  41. Shafer, W. M. and J. J. Iandolo. 1977. Chromosomal locus for staphylococcal enterotoxin B. J. Bacteriol. 129: 317- 325
  42. Shalita, Z., I. Hertman, and S. Sarid. 1985. Isolation and characterization of a plasmid involved with enterotoxin B production in Staphylococcus aureus. Infect. Immun. 47: 514-521
  43. Stiles, B. G. and K. Teresa. 2005. Staphylococcal enterotoxins: A purging experience in review, part I. Clin. Microbiol. Newslett. 27: 179-186 https://doi.org/10.1016/j.clinmicnews.2005.11.001
  44. Tamarapu, S., J. L. McKillip, and M. Drake. 2001. Development of a multiplex polymerase chain reaction assay for detection and differentiation of Staphylococcus aureus in dairy products. J. Food Prot. 64: 664-668 https://doi.org/10.4315/0362-028X-64.5.664
  45. Tremaine, M. T., D. K. Brockman, and M. J. Betley. 1993. Staphylococcal enterotoxin A gene (sea) expression is not affected by the accessory gene regulator (agr). Infect. Immun. 61: 356-359