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SDS-PAGE and Gel IEF: Tool for Differentiation of Methicillin-Resistant and Methicillin-Sensitive Strains of Staphylococcus aureus

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Abstract

The methicillin-resistant Staphylococcus aureus causes difficult-to-treat healthcare-associated infections in humans. For fast and effective selection of an appropriate antibiotic therapy, it is essential to have rapid and reliable methods for differentiation of methicillin-resistant S. aureus from less dangerous methicillin-sensitive S. aureus. There have been many methods for the identification of methicillin-resistant S. aureus described but none has been accepted as an international standard. The most commonly used techniques such as phenotyping and genotyping have a few disadvantages, for instance, these techniques are not reproducible and stable. In addition, they are time-consuming, expensive, and they are not capable to distinguish all S. aureus strains. In this study, the methicillin-resistant and methicillin-sensitive S. aureus isolates obtained from patients were extracted in hot water. The released proteins were characterised by sodium dodecyl sulphate polyacrylamide gel electrophoresis and gel isoelectric focusing. These two methods were able to differentiate among tested bacterial strains. The proposed methods are time saving, they are applicable in standard biochemical laboratories, and they do not require any expensive equipment.

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References

  1. AkonniBiosystems (2009) TruArray MRSA Test for Identifying MRSA and mobile mecA elements. http://www.akonni.com/docs/Akonni_TruArray_MRSA_Datasheet.pdf. Accessed 15 April 2015

  2. BioRad. Ready Gel, Precast gels, Application Guide, Catalog Number 151-0993. http://www.bio-rad.com/webroot/web/pdf/lsr/literature/LIT188H.pdf. Accessed 15 April 2015

  3. BioRad (2008) MRSA select, selective and differential chromogenic medium for the qualitative detection of nasal colonization of methicillin resistant Staphylococcus aureus (MRSA). Part No. 25285B. http://www.bio-rad.com/webroot/web/pdf/inserts/CDG/en/63747_08_2009_EN.pdf. Accessed 15 April 2015

  4. Costas M, Cookson BD, Talsania HG, Owen RJ (1989) Numerical analysis of electrophoretic protein patterns of methicillin-resistant strains of Staphylococcus aureus. J Clin Microbiol 27(11):2574–2581

    PubMed Central  CAS  PubMed  Google Scholar 

  5. DavisTestKits. http://www.daviskits.com/mrsa-test-kits/ Accessed 3 February 2015

  6. DENKA SEIKEN CO,. LTD. MRSA-screen a slide latex agglutination assay for the detection of PBP2 of S. aureus. 1K-06089B. http://denka-seiken.jp/english/pdf/mrsa-screen.pdf. Accessed 15 April 2015

  7. Doǧan G, Doǧanli GA, Gürsoy Y, Doǧan NM (2013) Antibiotic susceptibilities and SDS-PAGE protein profiles of methicillin-resistant Staphylococcus aureus (MRSA) strains obtained from Denizli Hospital. InTech. doi:10.5772/55457

    Google Scholar 

  8. European Centre for Disease Prevention and Control (2013) Annual epidemiological report Reporting on 2011 surveillance data and 2012 epidemic intelligence data. http://www.ecdc.europa.eu/en/publications/publications/annual-epidemiological-report-2013.pdf. Accessed 15 April 2015

  9. HAIN Lifescience. GenoQuick MRSA-When speed counts. http://www.hain-lifescience.de/en/products/microbiology/mrsa/genoquick-mrsa.html. Accessed 15 April 2015

  10. Horká M, Horký J, Matousková H, Slais K (2009) Free flow and capillary isoelectric focusing of bacteria from the tomatoes plant tissues. J Chromatogr A 1216(6):1019–1024. doi:10.1016/j.chroma.2008.12.024

    Article  PubMed  Google Scholar 

  11. Horká M, Karásek P, Salplachta J, Růžička F, Vykydalová M, Kubesová A, Dráb V, Roth M, Slais K (2013) Capillary isoelectric focusing of probiotic bacteria from cow’s milk in tapered fused silica capillary with off-line matrix-assisted laser desorption/ionization time-of-flight mass spectrometry identification. Anal Chim Acta 788:193–199. doi:10.1016/j.aca.2013.05.059

    Article  PubMed  Google Scholar 

  12. Horká M, Tesařová M, Karásek P, Růžička F, Holá V, Sittová M, Roth M (2015) Determination of methicillin-resistant and methicillin-susceptible Staphylococcus aureus bacteria in blood by capillary zone electrophoresis. Anal Chim Acta 868:62–72. doi:10.1016/j.aca.2015.02.00113

    Article  Google Scholar 

  13. Hussein MY, Gohar ZM (2006) A new approach for MRSA blind chemotherapy. Egypt J Med Microbiol 15(1):1–16

    Google Scholar 

  14. Kohrs F, Heyer R, Magnussen A, Benndorf D, Muth T, Behne A, Rapp E, Kausmann R, Heiermann M, Klocke M, Reichl U (2014) Sample prefractionation with liquid isoelectric focusing enables in depth microbial metaproteome analysis of mesophilic and thermophilic biogas plants. Anaerobe 29:59–67. doi:10.1016/j.anaerobe.2013.11.009

    Article  CAS  PubMed  Google Scholar 

  15. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227(5259):680–685

    Article  CAS  PubMed  Google Scholar 

  16. Liu Z, Wu SS, Pawliszyn J (2007) Characterization of plant growth-promoting rhizobacteria using capillary isoelectric focusing with whole column imaging detection. J Chromatogr A 1140(1–2):213–218

    Article  CAS  PubMed  Google Scholar 

  17. Otto M (2010) Basis of virulence in community-associated methicillin-resistant Staphylococcus aureus. Annu Rev Microbiol 64:143–162. doi:10.1093/infdis/jiu462

    Article  CAS  PubMed  Google Scholar 

  18. Rajaduraipandi K, Panneerselvam K, Ravikumar K, Rajasekaran P, Manoharan C, Shanmugam S (2008) Typing of methicillin resistant Staphylococcus aureus using whole cell polypeptide and immunoblotting techniques. Adv Biotech 6:14–17

    Google Scholar 

  19. Ravipaty S, Reilly JP (2010) Comprehensive characterization of methicillin-resistant Staphylococcus aureus subsp. aureus COL secretome by two-dimensional liquid chromatography and mass spectrometry. Mol Cell Proteom 9(9):1898–1919. doi:10.1074/mcp.M900494-MCP200

    Article  CAS  Google Scholar 

  20. Ruzicka F, Horka M, Hola V, Votava M (2007) Capillary isoelectric focusing—useful tool for detection of the biofilm formation in Staphylococcus epidermidis. J Microbiol Methods 68(3):530–535

    Article  CAS  PubMed  Google Scholar 

  21. Saҫilik SC, Osmanaǧaoǧlu Ö, Kisa Ö, Başustaoǧlu A, Çökmüş C (2000) Protein profiles and prevalence of methicillin resistant Staphylococcus aureus (MRSA) in Gülhane Military Academy Hospital in Turkey. Turk J Biol 24:809–816

    Google Scholar 

  22. Saҫilik SC, Osmanaǧaoǧlu Ö, Palabiyikoǧlu I, Bengisun JS, Çökmüş C (2000) Analysis methicillin resistant Staphylococcus aureus isolates by polyacrylamide gel electrophoresis in an intensive care unit of İbni-Sina Hospital. Turk J Biol 30:367–671

    Google Scholar 

  23. Schägger H, Von Jagow G (1987) Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Anal Biochem 166(2):368–379

    Article  PubMed  Google Scholar 

  24. Thomson-Carter FM, Pennington TH (1989) Characterization of coagulase-negative staphylococci by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblot analyses. Clin Microbiol 27(10):2199–2203

    CAS  Google Scholar 

  25. Torimiro N, Ako-Nai AK, Omitogun OG (2013) Comparative protein profiling of antibiotic resistant S. aureus strains from hospital and community-associated infections. Int Res J Microbiol 4(2):64–71

    Google Scholar 

  26. Vykydalová M, Horká M, Růžička F, Duša F, Moravcová D, Kahle V (2014) Combination of micropreparative solution isoelectric focusing and high-performance liquid chromatography for differentiation of biofilm-positive and biofilm-negative Candida parapsilosis group from vascular catheter. Anal Chim Acta 812:243–249. doi:10.1016/j.aca.2013.12.032

    Article  PubMed  Google Scholar 

  27. Weller TM (2000) Methicillin-resistant Staphylococcus aureus typing methods: which should be the international standard? J Hosp Infect 44(3):160–172

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

This study has been supported by the Grant of the Ministry of the Interior of the Czech Republic No. VG20112015021, by the Internal Grant Agency of the Ministry of Health of the Czech Republic NT/13242-4, by INGO II (LG13011), and by received institutional support RVO:68081715.

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Authors and Affiliations

  1. Institute of Analytical Chemistry of the CAS, v. v. i., Veveří 97, 602 00, Brno, Czech Republic

    Marie Tesařová, Marie Horká, Dana Moravcová & Lenka Svojanovská

  2. The Department of Microbiology, Faculty of Medicine, Masaryk University, Kamenice 53/5, 625 00, Brno, Czech Republic

    Katarina Mlynarikova & Filip Růžička

  3. The Department of Microbiology, St. Anne’s University Hospital, Brno, Pekařská 53, 602 00, Brno, Czech Republic

    Filip Růžička

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  1. Marie Tesařová
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  2. Marie Horká
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Correspondence to Marie Tesařová.

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Tesařová, M., Horká, M., Moravcová, D. et al. SDS-PAGE and Gel IEF: Tool for Differentiation of Methicillin-Resistant and Methicillin-Sensitive Strains of Staphylococcus aureus . Curr Microbiol 72, 315–320 (2016). https://doi.org/10.1007/s00284-015-0939-y

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  • DOI: https://doi.org/10.1007/s00284-015-0939-y

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