spa Typing for Epidemiological Surveillance of Staphylococcus aureus

  • Marie Hallin
  • Alexander W. Friedrich
  • Marc J. Struelens
Protocol
Part of the Methods in Molecular Biology™ book series (MIMB, volume 551)

Abstract

The spa typing method is based on sequencing of the polymorphic X region of the protein A gene (spa), present in all strains of Staphylococcus aureus. The X region is constituted of a variable number of 24-bp repeats flanked by well-conserved regions. This single-locus sequence-based typing method combines a number of technical advantages, such as rapidity, reproducibility, and portability. Moreover, due to its repeat structure, the spa locus simultaneously indexes micro- and macrovariations, enabling the use of spa typing in both local and global epidemiological studies. These studies are facilitated by the establishment of standardized spa type nomenclature and Internet shared databases.

Key words

Epidemiology methicillin resistance phylogeny S. aureus sequence analysis software staphylococcal protein A typing methods 

References

  1. 1.
    Lowy, F. D. (1998). Staphylococcus aureus infections. N. Engl. J. Med. 339, 520–532.PubMedCrossRefGoogle Scholar
  2. 2.
    Jevons, M. P. (1961). “Celbenin”-resistant Staphylococci. BMJ 1, 124–125.CrossRefGoogle Scholar
  3. 3.
    Aires, d. S., and de Lencastre, H. (2004). Bridges from hospitals to the laboratory: genetic portraits of methicillin-resistant Staphylococcus aureus clones. FEMS Immunol. Med. Microbiol. 40, 101–111.CrossRefGoogle Scholar
  4. 4.
    Strommenger, B., Kettlitz, C., Weniger, T., Harmsen, D., Friedrich, A. W., and Witte, W. (2006). Assignment of Staphylococcus isolates to groups by spa typing, SmaI macrorestriction analysis, and multilocus sequence typing. J. Clin. Microbiol. 44, 2533–2540.PubMedCrossRefGoogle Scholar
  5. 5.
    Tenover, F. C., Arbeit, R., Archer, G., Biddle, J., Byrne, S., Goering, R., et al (1994). Comparison of traditional and molecular methods of typing isolates of Staphylococcus aureus. J. Clin. Microbiol. 32, 407–415.PubMedGoogle Scholar
  6. 6.
    Murchan, S., Kaufmann, M. E., Deplano, A., De Ryck, R., Struelens, M., Zinn, C. E., et al (2003). Harmonization of pulsed-field gel electrophoresis protocols for epidemiological typing of strains of methicillin-resistant Staphylococcus aureus: a single approach developed by consensus in 10 European laboratories and its application for tracing the spread of related strains. J. Clin. Microbiol. 41, 1574–1585.PubMedCrossRefGoogle Scholar
  7. 7.
    Deplano, A., Schuermans, A., Van Eldere, J., Witte, W., Meugnier, H., Etienne, J., et al (2000). Multicenter evaluation of epidemiological typing of methicillin-resistant Staphylococcus aureus strains by repetitive-element PCR analysis. The European Study Group on Epidemiological Markers of the ESCMID.J. Clin. Microbiol. 38, 3527–3533.PubMedGoogle Scholar
  8. 8.
    Cookson, B. D., Aparicio, P., Deplano, A., Struelens, M., Goering, R., and Marples, R. (1996). Inter-centre comparison of pulsed-field gel electrophoresis for the typing of methicillin-resistant Staphylococcus aureus. J. Med. Microbiol. 44, 179–184.PubMedCrossRefGoogle Scholar
  9. 9.
    Enright, M. C., Day, N. P., Davies, C. E., Peacock, S. J., and Spratt, B. G. (2000). Multilocus sequence typing for characterization of methicillin-resistant and methicillin-susceptible clones of Staphylococcus aureus. J. Clin. Microbiol. 38, 1008–1015.PubMedGoogle Scholar
  10. 10.
    Enright, M. C., Robinson, D. A., Randle, G., Feil, E. J., Grundmann, H., and Spratt, B. G. (2002). The evolutionary history of methicillin-resistant Staphylococcus aureus (MRSA). Proc. Natl. Acad. Sci. U S A 99, 7687–7692.PubMedCrossRefGoogle Scholar
  11. 11.
    Peacock, S. J., de Silva, G. D., Justice, A., Cowland, A., Moore, C. E., Winearls, C. G., et al (2002). Comparison of multilocus sequence typing and pulsed-field gel electrophoresis as tools for typing Staphylococcus aureus isolates in a microepidemiological setting. J. Clin. Microbiol. 40, 3764–3770.PubMedCrossRefGoogle Scholar
  12. 12.
    Frenay, H. M., Theelen, J. P., Schouls, L. M., Vandenbroucke-Grauls, C. M., Verhoef, J., van Leeuwen, W. J., et al (1994). Discrimination of epidemic and nonepidemic methicillin-resistant Staphylococcus aureus strains on the basis of protein A gene polymorphism. J. Clin. Microbiol. 32, 846–847.PubMedGoogle Scholar
  13. 13.
    Frenay, H. M., Bunschoten, A. E., Schouls, L. M., van Leeuwen, W. J., Vandenbroucke-Grauls, C. M., Verhoef, J., et al (1996). Molecular typing of methicillin-resistant Staphylococcus aureus on the basis of protein A gene polymorphism. Eur. J. Clin. Microbiol. Infect. Dis. 15, 60–64.PubMedCrossRefGoogle Scholar
  14. 14.
    Guss, B., Uhlen, M., Nilsson, B., Lindberg, M., Sjoquist, J., and Sjodahl, J. (1984). Region X, the cell-wall-attachment part of staphylococcal protein A. Eur. J. Biochem. 138, 413–420.PubMedCrossRefGoogle Scholar
  15. 15.
    van Belkum, A. (1999). The role of short sequence repeats in epidemiologic typing. Curr. Opin. Microbiol. 2, 306–311.PubMedCrossRefGoogle Scholar
  16. 16.
    van Belkum, A. (2007). Tracing isolates of bacterial species by multilocus variable number of tandem repeat analysis (MLVA). FEMS Immunol. Med. Microbiol. 49, 22–27.PubMedCrossRefGoogle Scholar
  17. 17.
    Harmsen, D., Claus, H., Witte, W., Rothganger, J., Claus, H., Turnwald, D., et al (2003). Typing of methicillin-resistant Staphylococcus aureus in a university hospital setting by using novel software for spa repeat determination and database management. J. Clin. Microbiol. 41, 5442–5448.PubMedCrossRefGoogle Scholar
  18. 18.
    Shopsin, B., Gomez, M., Montgomery, S. O., Smith, D. H., Waddington, M., Dodge, D. E., et al (1999). Evaluation of protein A gene polymorphic region DNA sequencing for typing of Staphylococcus aureus strains. J. Clin. Microbiol. 37, 3556–3563.PubMedGoogle Scholar
  19. 19.
    van Belkum, A., Tassios, P. T., Dijkshoorn, L., Haeggman, S., Cookson, B., Fry, N. K., et al (2007). Guidelines for the validation and application of typing methods for use in bacterial epidemiology. Clin. Microbiol. Infect. 13 (Suppl. 3), 1–46.PubMedCrossRefGoogle Scholar
  20. 20.
    Koreen, L., Ramaswamy, S. V., Graviss, E. A., Naidich, S., Musser, J. M., and Kreiswirth, B. N. (2004). spa typing method for discriminating among Staphylococcus aureus isolates: implications for use of a single marker to detect genetic micro- and macrovariation. J. Clin. Microbiol. 42, 792–799.PubMedCrossRefGoogle Scholar
  21. 21.
    Aires-de-Sousa, M., Boye, K., de Lencastre, H., Deplano, A., Enright, M. C., Etienne, J., et al (2006). High interlaboratory reproducibility of DNA sequence-based typing of bacteria in a multicenter study. J. Clin. Microbiol. 44, 619–621.PubMedCrossRefGoogle Scholar
  22. 22.
    Kahl, B. C., Mellmann, A., Deiwick, S., Peters, G., and Harmsen, D. (2005). Variation of the polymorphic region X of the protein A gene during persistent airway infection of cystic fibrosis patients reflects two independent mechanisms of genetic change in Staphylococcus aureus. J. Clin. Microbiol. 43, 502–505.PubMedCrossRefGoogle Scholar
  23. 23.
    Goerke, C., Gressinger, M., Endler, K., Breitkopf, C., Wardecki, K., Stern, M., et al (2007). High phenotypic diversity in infecting but not in colonizing Staphylococcus aureus populations. Environ. Microbiol. 9, 3134–3142.PubMedCrossRefGoogle Scholar
  24. 24.
    Hallin, M., Deplano, A., Denis, O., de Mendonca, R., De Ryck, R., and Struelens, M. J. (2007). Validation of pulsed-field gel electrophoresis and spa typing for long-term, nationwide epidemiological surveillance studies of Staphylococcus aureus infections. J. Clin. Microbiol. 45, 127–133.PubMedCrossRefGoogle Scholar
  25. 25.
    Tenover, F. C., Arbeit, R. D., Goering, R. V., Mickelsen, P. A., Murray, B. E., Persing, D. H., et al (1995). Interpreting chromosomal DNA restriction patterns produced by pulsed-field gel electrophoresis: criteria for bacterial strain typing. J. Clin. Microbiol. 33, 2233–2239.PubMedGoogle Scholar
  26. 26.
    Faria, N. A., Carrico, J. A., Oliveira, D. C., Ramirez, M., and de Lencastre, H. (2008). Analysis of typing methods for epidemiological surveillance of both methicillin-resistant and methicillin-susceptible Staphylococcus aureus strains. J. Clin. Microbiol. 46, 136–144.PubMedCrossRefGoogle Scholar
  27. 27.
    Strommenger, B., Braulke, C., Heuck, D., Schmidt, C., Pasemann, B., Nubel, U., et al (2008). spa typing of Staphylococcus aureus as a frontline tool in epidemiological typing J. Clin. Microbiol. 46, 574–581.PubMedCrossRefGoogle Scholar
  28. 28.
    Robinson, D. A., and Enright, M. C. (2004). Evolution of Staphylococcus aureus by large chromosomal replacementsJ. Bacteriol. 186, 1060–1064.PubMedCrossRefGoogle Scholar
  29. 29.
    Ruppitsch, W., Indra, A., Stoger, A., Mayer, B., Stadlbauer, S., Wewalka, G., et al (2006). Classifying spa types in complexes improves interpretation of typing results for methicillin-resistant Staphylococcus aureus. J. Clin. Microbiol. 44, 2442–2448.PubMedCrossRefGoogle Scholar
  30. 30.
    Mellmann, A., Friedrich, A. W., Rosenkotter, N., Rothganger, J., Karch, H., Reintjes, R., et al (2006). Automated DNA sequence-based early warning system for the detection of methicillin-resistant Staphylococcus aureus outbreaks. PLoS Med. 3, e33.PubMedCrossRefGoogle Scholar
  31. 31.
    Oliveira, D. C., Crisostomo, I., Santos-Sanches, I., Major, P., Alves, C. R., Aires-de-Sousa, M., et al (2001). Comparison of DNA sequencing of the protein A gene polymorphic region with other molecular typing techniques for typing two epidemiologically diverse collections of methicillin-resistant Staphylococcus aureus. J. Clin. Microbiol. 39, 574–580.PubMedCrossRefGoogle Scholar
  32. 32.
    Feil, E. J., Cooper, J. E., Grundmann, H., Robinson, D. A., Enright, M. C., Berendt, T., et al (2003). How clonal is Staphylococcus aureus? J. Bacteriol. 185, 3307–3316.CrossRefGoogle Scholar
  33. 33.
    Robinson, D. A., and Enright, M. C. (2003). Evolutionary models of the emergence of methicillin-resistant Staphylococcus aureus. Antimicrob. Agents Chemother. 47, 3926–3934.PubMedCrossRefGoogle Scholar
  34. 34.
    Mellmann, A., Weniger, T., Berssenbrugge, C., Rothganger, J., Sammeth, M., Stoye, J., et al (2007). Based Upon Repeat Pattern (BURP): an algorithm to characterize the long-term evolution of Staphylococcus aureus populations based on spa polymorphisms. BMC Microbiol. 7, 98.PubMedCrossRefGoogle Scholar
  35. 35.
    Friedrich, A. W., Witte, W., Harmsen, D., de Lencastre, H., Hryniewicz, W., Scheres, J., et al (2006). SeqNet.org: a European laboratory network for sequence-based typing of microbial pathogens. Euro Surveill. 11, E060112.PubMedGoogle Scholar
  36. 36.
    von Eiff, C., Maas, D., Sander, G., Friedrich, A. W., Peters, G., and Becker, K. (2008). Microbiological evaluation of a new growth-based approach for rapid detection of methicillin-resistant Staphylococcus aureus. J. Antimicrob. Chemother. 61, 1277–1280.PubMedCrossRefGoogle Scholar
  37. 37.
    Friedrich, A., Daniels-Haardt, I., Gemert-Pijnen, J., Hendrix, M., von Eiff, C., Becker, K., et al (2007). A regional network for the prevention and control of infections due to MRSA: EUREGIO MRSA-net Twente/Münsterland. Epidemiol. Bull. 33, 307–311.Google Scholar
  38. 38.
    Daniels-Haardt, I., Verhoeven, F., Mellmann, A., Hendrix, M. G., Gemert-Pijnen, J. E., and Friedrich, A. W. (2006). [EUREGIO-projekt MRSA-net Twente/Munsterland. Creation of a regional network to combat MRSA]. Gesundheitswesen 68, 674–678.PubMedCrossRefGoogle Scholar
  39. 39.
    Deurenberg, R. H., Vink, C., Oudhuis, G. J., Mooij, J. E., Driessen, C., Coppens, G., et al (2005). Different clonal complexes of methicillin-resistant Staphylococcus aureus are disseminated in the Euregio Meuse-Rhine region. Antimicrob. Agents Chemother. 49, 4263–4271.PubMedCrossRefGoogle Scholar
  40. 40.
    Oliveira, D. C., Crisostomo, I., Santos-Sanches, I., Major, P., Alves, C. R., Aires-de-Sousa, M., et al (2001). Comparison of DNA sequencing of the protein A gene polymorphic region with other molecular typing techniques for typing two epidemiologically diverse collections of methicillin-resistant Staphylococcus aureus. J. Clin. Microbiol. 39, 574–580.PubMedCrossRefGoogle Scholar
  41. 41.
    Uhlen, M., Guss, B., Nilsson, B., Gatenbeck, S., Philipson, L., and Lindberg, M. (1984). Complete sequence of the staphylococcal gene encoding protein A. A gene evolved through multiple duplications. J. Biol. Chem. 259, 1695–1702.PubMedGoogle Scholar

Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Marie Hallin
    • 1
  • Alexander W. Friedrich
    • 1
  • Marc J. Struelens
    • 1
  1. 1.Laboratoire de Référence MRSA-StaphylocoquesDepartment of Microbiology, Hôpital Erasme, Université Libre de BruxellesBrusselsBelgium

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