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Staphylococcal Cassette Chromosome mec (SCCmec) Analysis of MRSA

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Methicillin-Resistant Staphylococcus Aureus (MRSA) Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1085))

Abstract

Methicillin-susceptible S. aureus (MSSA) changes to methicillin-resistant S. aureus upon the acquisition of Staphylococcal Cassette Chromosome mec (SCCmec), a genomic island that encodes methicillin resistance. All SCCmec elements reported to date share four common characteristics: (1) carrying the mec gene complex (mec); (2) carrying the ccr gene complex (ccr); (3) being flanked by characteristic nucleotide sequences, inverted repeats, and direct repeats, at both ends; and (4) being integrated at the integration site sequence (ISS) for SCC, which is located at the 3′-end of orfX or at the extremity of the SCC element. SCCmec elements in S. aureus are classified into different types based on the combination of mec and ccr, which share variations, five classes in mec and eight in ccr. To date, at least 11 types of SCCmec elements have been identified. Regions other than mec and ccr within the SCCmec element are designated as “joining regions” (J-regions), which are classified into three subgroups, J1-3. Many J-region variants have been identified among the SCCmec elements of types I–V. We herein describe PCR methods to type SCCmec elements by first identifying the mec and ccr type, and then identifying genes in the J-regions.

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References

  1. Ito T, Katayama Y, Hiramatsu K (1999) Cloning and nucleotide sequence determination of the entire mec DNA of pre-methicillin-resistant Staphylococcus aureus N315. Antimicrob Agents Chemother 43:1449–1458

    PubMed  CAS  Google Scholar 

  2. Katayama Y, Ito T, Hiramatsu K (2000) A new class of genetic element, staphylococcal cassette chromosome mec, encodes methicillin resistance in Staphylococcus aureus. Antimicrob Agents Chemother 44:1549–1555

    Article  PubMed  CAS  Google Scholar 

  3. International Working Group on the Classification of Staphylococcal Cassette Chromosome Elements (IWG-SCC) (2009) Classification of staphylococcal cassette chromosome mec (SCCmec): guidelines for reporting novel SCCmec elements. Antimicrob Agents Chemother 53:4961–4967

    Article  Google Scholar 

  4. Ito T, Katayama Y, Asada K et al (2001) Structural comparison of three types of staphylococcal cassette chromosome mec integrated in the chromosome in methicillin-resistant Staphylococcus aureus. Antimicrob Agents Chemother 45:1323–1336

    Article  PubMed  CAS  Google Scholar 

  5. Ma XX, Ito T, Tiensasitorn C et al (2002) Novel type of staphylococcal cassette chromosome mec identified in community-acquired methicillin-resistant Staphylococcus aureus strains. Antimicrob Agents Chemother 46:1147–1152

    Article  PubMed  CAS  Google Scholar 

  6. Ito T, Ma XX, Takeuchi F et al (2004) Novel type V staphylococcal cassette chromosome mec driven by a novel cassette chromosome recombinase, ccrC. Antimicrob Agents Chemother 48:2637–2651

    Article  PubMed  CAS  Google Scholar 

  7. Oliveira DC, Milheirico C, de Lencastre H (2006) Redefining a structural variant of staphylococcal cassette chromosome mec, SCCmec type VI. Antimicrob Agents Chemother 50:3457–3459

    Article  PubMed  CAS  Google Scholar 

  8. Berglund C, Ito T, Ikeda M et al (2008) Novel type of staphylococcal cassette chromosome mec in a methicillin-resistant Staphylococcus aureus strain isolated in Sweden. Antimicrob Agents Chemother 52:3512–3516

    Article  PubMed  CAS  Google Scholar 

  9. Zhang K, McClure JA, Elsayed S et al (2009) Novel staphylococcal cassette chromosome mec type, tentatively designated type VIII, harboring class A mec and type 4 ccr gene complexes in a Canadian epidemic strain of methicillin-resistant Staphylococcus aureus. Antimicrob Agents Chemother 53:531–540

    Article  PubMed  CAS  Google Scholar 

  10. Li S, Skov RL, Han X et al (2011) Novel types of staphylococcal cassette chromosome mec elements identified in clonal complex 398 methicillin-resistant Staphylococcus aureus strains. Antimicrob Agents Chemother 55:3046–3050

    Article  PubMed  CAS  Google Scholar 

  11. Garcia-Alvarez L, Holden MT, Lindsay H et al (2011) Meticillin-resistant Staphylococcus aureus with a novel mecA homologue in human and bovine populations in the UK and Denmark: a descriptive study. Lancet Infect Dis 11:595–603

    Article  PubMed  CAS  Google Scholar 

  12. Katayama Y, Ito T, Hiramatsu K (2001) Genetic organization of the chromosome region surrounding mecA in clinical Staphylococcal strains: role of IS431-mediated mecI deletion in expression of resistance in mecA-carrying, low-level methicillin- resistant Staphylococcus haemolyticus. Antimicrob Agents Chemother 45:1955–1963

    Article  PubMed  CAS  Google Scholar 

  13. Ryffel C, Bucher R, Kayser FH et al (1991) The Staphylococcus aureus mec determinant comprises an unusual cluster of direct repeats and codes for gene product similar to the Escherichia coli sn-glycerophosphoryl diester phosphodiesterase. J Bacteriol 173:7416–7422

    PubMed  CAS  Google Scholar 

  14. Goering RV, Morrison D, Al-Doori Z et al (2008) Usefulness of mec-associated direct repeat unit (dru) typing in the epidemiological analysis of highly clonal methicillin-resistant Staphylococcus aureus in Scotland. Clin Microbiol Infect 14:964–969

    Article  PubMed  CAS  Google Scholar 

  15. Ito T, Hiramatsu K, Tomasz A et al (2012) Guidelines for reporting novel mecA gene homologues. Antimicrob Agents Chemother 56:4997–4999

    Article  PubMed  CAS  Google Scholar 

  16. Chambers HF (2001) The changing epidemiology of Staphylococcus aureus. Emerg Infect Dis 7:178–182

    Article  PubMed  CAS  Google Scholar 

  17. Eady EA, Cove JH (2003) Staphylococcal resistance revisited: community-acquired methicillin resistant Staphylococcus aureus—an emerging problem for the management of skin and soft tissue infections. Curr Opin Infect Dis 16:103–124

    Article  PubMed  CAS  Google Scholar 

  18. Okuma K, Iwakawa K, Turnidge JD et al (2002) Dissemination of new methicillin-resistant Staphylococcus aureus clones in the community. J Clin Microbiol 40:4289–4294

    Article  PubMed  CAS  Google Scholar 

  19. Chambers HF, Deleo FR (2009) Waves of resistance: Staphylococcus aureus in the antibiotic era. Nat Rev Microbiol 7:629–641

    Article  PubMed  CAS  Google Scholar 

  20. Schijffelen MJBC, van Strijp JA, Fluit AC (2010) Whole genome analysis of a livestock-associated methicillin-resistant Staphylococcus aureus ST398 isolate from a case of human endocarditis. BMC Genomics 11:376

    Article  PubMed  Google Scholar 

  21. Shore AC, Deasy EC, Slickers P et al (2011) Detection of Staphylococcal cassette chromosome mec type XI carrying highly divergent mecA, mecI, mecR1, blaZ, and ccr genes in human clinical isolates of clonal complex 130 methicillin-resistant Staphylococcus aureus. Antimicrob Agents Chemother 55:3765–3773

    Article  PubMed  CAS  Google Scholar 

  22. Han X, Ito T, Takeuchi F et al (2009) Identification of a novel variant of staphylococcal cassette chromosome mec, type II.5, and its truncated form by insertion of putative conjugative transposon Tn6012. Antimicrob Agents Chemother 53:2616–2619

    Article  PubMed  CAS  Google Scholar 

  23. Kondo Y, Ito T, Ma XX et al (2007) Combination of multiplex PCRs for staphylococcal cassette chromosome mec type assignment: rapid identification system for mec, ccr, and major differences in junkyard regions. Antimicrob Agents Chemother 51:264–274

    Article  PubMed  CAS  Google Scholar 

  24. Ma XX, Ito T, Chongtrakool P, Hiramatsu K (2006) Predominance of clones carrying Panton-Valentine leukocidin genes among methicillin-resistant Staphylococcus aureus strains isolated in Japanese hospitals from 1979 to 1985. J Clin Microbiol 44:4515–4527

    Article  PubMed  CAS  Google Scholar 

  25. Berglund C, Ito T, Ma XX et al (2009) Genetic diversity of methicillin-resistant Staphylococcus aureus carrying type IV SCCmec in Orebro County and the western region of Sweden. J Antimicrob Chemother 63:32–41

    Article  PubMed  CAS  Google Scholar 

  26. Kwon NH, Park KT, Moon JS et al (2005) Staphylococcal cassette chromosome mec (SCCmec) characterization and molecular analysis for methicillin-resistant Staphylococcus aureus and novel SCCmec subtype IVg isolated from bovine milk in Korea. J Antimicrob Chemother 56:624–632

    Article  PubMed  CAS  Google Scholar 

  27. Milheirico C, Oliveira DC, de Lencastre H (2007) Multiplex PCR strategy for subtyping the staphylococcal cassette chromosome mec type IV in methicillin-resistant Staphylococcus aureus: ‘SCCmec IV multiplex’. J Antimicrob Chemother 60:42–48

    Article  PubMed  CAS  Google Scholar 

  28. Cavaco LM, Hasman H, Stegger M et al (2010) Cloning and occurrence of czrC, a gene conferring cadmium and zinc resistance in methicillin-resistant Staphylococcus aureus CC398 isolates. Antimicrob Agents Chemother 54:3605–3608

    Article  PubMed  CAS  Google Scholar 

  29. Oliveira DC, Lencastre H (2002) Multiplex PCR strategy for rapid identification of structural types and variants of the mec element in methicillin-resistant Staphylococcus aureus. Antimicrob Agents Chemother 46:2155–2161

    Article  PubMed  CAS  Google Scholar 

  30. Zhang K, McClure JA, Elsayed S et al (2005) Novel multiplex PCR assay for characterization and concomitant subtyping of staphylococcal cassette chromosome mec types I to V in methicillin-resistant Staphylococcus aureus. J Clin Microbiol 43:5026–5033

    Article  PubMed  CAS  Google Scholar 

  31. Zhang K, McClure JA, Conly JM (2012) Enhanced multiplex PCR assay for typing of staphylococcal cassette chromosome mec types I to V in methicillin-resistant Staphylococcus aureus. Mol Cell Probes 26:218–221

    Article  PubMed  Google Scholar 

  32. Milheirico C, Oliveira DC, de Lencastre H (2007) Update to the multiplex PCR strategy for assignment of mec element types in Staphylococcus aureus. Antimicrob Agents Chemother 51:3374–3377

    Article  PubMed  CAS  Google Scholar 

  33. Oliveira DC, Wu SW, Lencastre H (2000) Genetic organization of the downstream region of the mecA element in methicillin-resistant Staphylococcus aureus isolates carrying different polymorphisms of this region. Antimicrob Agents Chemother 44:1906–1910

    Article  PubMed  CAS  Google Scholar 

  34. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning. Cold Spring Harbor Laboratory Press, USA, New York

    Article  PubMed  CAS  Google Scholar 

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Acknowledgement

This work was supported by a Grant-in-Aid from MEXT (Ministry of Education, Culture, Sports, Science and Technology)—Supported Program for the Strategic Research Foundation at Private Universities.

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Authors and Affiliations

  1. Department of Infection Control Science, Graduate School of Medicine, Juntendo University, Tokyo, Japan

    Teruyo Ito, Kyoko Kuwahara-Arai, Yuki Katayama, Yuki Uehara, Xiao Han, Yoko Kondo & Keiichi Hiramatsu

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  1. Teruyo Ito
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  2. Kyoko Kuwahara-Arai
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  3. Yuki Katayama
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  4. Yuki Uehara
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  5. Xiao Han
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  6. Yoko Kondo
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  7. Keiichi Hiramatsu
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Editors and Affiliations

  1. Dept. Veterinary & Biomedical Sciences, University of Minnesota College of Veterinary Medicine, St. Paul, Minnesota, USA

    Yinduo Ji

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Ito, T. et al. (2014). Staphylococcal Cassette Chromosome mec (SCCmec) Analysis of MRSA. In: Ji, Y. (eds) Methicillin-Resistant Staphylococcus Aureus (MRSA) Protocols. Methods in Molecular Biology, vol 1085. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-664-1_8

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  • DOI: https://doi.org/10.1007/978-1-62703-664-1_8

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-663-4

  • Online ISBN: 978-1-62703-664-1

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