Skip to main content

Advertisement

SpringerLink
Log in

Rapid detection of methicillin-resistant staphylococci by real-time PCR directly from positive blood culture bottles

  • Brief Report
  • Published:
European Journal of Clinical Microbiology & Infectious Diseases Aims and scope Submit manuscript

Abstract

For the rapid detection of methicillin-resistant staphylococci directly from blood cultures containing gram-positive cocci in clusters, we implemented a real-time (LightCycler) polymerase chain reaction (PCR) specific for the Staphylococcus aureus nuc gene encoding nuclease and the mecA gene encoding methicillin resistance. For the 475 positive blood cultures tested, the assay turned out to have 100% sensitivity and 100% specificity for the identification of methicillin-susceptible (n = 108) and methicillin-resistant (n = 34) S. aureus. When coagulase-negative staphylococci (CoNS) were included, the overall sensitivity for the detection of methicillin resistance was 93% and the specificity was 99%. Real-time PCR for nuc and mecA from blood culture bottles with staphylococci yields therefore a rapid (2–3 h) identification of S. aureus and CoNS including methicillin resistance.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

References

  1. Annane D, Bellissant E, Cavaillon JM (2005) Septic shock. Lancet 365:63–78

    Article  PubMed  CAS  Google Scholar 

  2. Cosgrove SE, Sakoulas G, Perencevich EN, Schwaber MJ, Karchmer AW, Carmeli Y (2003) Comparison of mortality associated with methicillin-resistant and methicillin-susceptible Staphylococcus aureus bacteremia: a meta-analysis. Clin Infect Dis 36:53–59

    Article  PubMed  Google Scholar 

  3. Beekmann SE, Diekema DJ, Chapin KC, Doern GV (2003) Effects of rapid detection of bloodstream infections on length of hospitalization and hospital charges. J Clin Microbiol 41:3119–3125

    Article  PubMed  CAS  Google Scholar 

  4. MacGowan AP (1998) Pharmacodynamics, pharmacokinetics, and therapeutic drug monitoring of glycopeptides. Ther Drug Monit 20:473–477

    Article  PubMed  CAS  Google Scholar 

  5. Maor Y, Rahav G, Belausov N, Ben-David D, Smollan G, Keller N (2007) Prevalence and characteristics of heteroresistant vancomycin-intermediate Staphylococcus aureus (hVISA) bacteremia in a tertiary care center. J Clin Microbiol 45:1511–1514

    Article  PubMed  CAS  Google Scholar 

  6. Small PM, Chambers HF (1990) Vancomycin for Staphylococcus aureus endocarditis in intravenous drug users. Antimicrob Agents Chemother 34:1227–1231

    PubMed  CAS  Google Scholar 

  7. Reischl U, Linde HJ, Metz M, Leppmeier B, Lehn N (2000) Rapid identification of methicillin-resistant Staphylococcus aureus and simultaneous species confirmation using real-time fluorescence PCR. J Clin Microbiol 38:2429–2433

    PubMed  CAS  Google Scholar 

  8. Clinical and Laboratory Standards Institute (2005) Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically. Approved standard M7-A7. Clinical and Laboratory Standards Institute, Wayne, PA

    Google Scholar 

  9. Clinical and Laboratory Standards Institute (2005) Performance standards for antimicrobial disk susceptibility tests. Approved standard M2-A9. Clinical and Laboratory Standards Institute, Wayne, PA

    Google Scholar 

  10. Clinical and Laboratory Standards Institute (2005) Performance standards for antimicrobial susceptibility testing. 15th informational supplement (M100-S15). Clinical and Laboratory Standards Institute, Wayne, PA

    Google Scholar 

  11. Sakai H, Procop GW, Kobayashi N, Togawa D, Wilson DA, Borden L, Krebs V, Bauer TW (2004) Simultaneous detection of Staphylococcus aureus and coagulase-negative staphylococci in positive blood cultures by real-time PCR with two fluorescence resonance energy transfer probe sets. J Clin Microbiol 42:5739–5744

    Article  PubMed  CAS  Google Scholar 

  12. Paule SM, Pasquariello AC, Thomson RB Jr, Kaul KL, Peterson LR (2005) Real-time PCR can rapidly detect methicillin-susceptible and methicillin-resistant Staphylococcus aureus directly from positive blood culture bottles. Am J Clin Pathol 124:404–407

    Article  PubMed  CAS  Google Scholar 

  13. Knobloch JK, Jäger S, Huck J, Horstkotte MA, Mack D (2005) mecA is not involved in the sigmaB-dependent switch of the expression phenotype of methicillin resistance in Staphylococcus epidermidis. Antimicrob Agents Chemother 49:1216–1219

    Article  PubMed  CAS  Google Scholar 

  14. Horstkotte MA, Knobloch JK, Rohde H, Dobinsky S, Mack D (2004) Evaluation of the BD PHOENIX automated microbiology system for detection of methicillin resistance in coagulase-negative staphylococci. J Clin Microbiol 42:5041–5046

    Article  PubMed  CAS  Google Scholar 

  15. Liu H, Buescher G, Lewis N, Snyder S, Jungkind D (1990) Detection of borderline oxacillin-resistant Staphylococcus aureus and differentiation from methicillin-resistant strains. Eur J Clin Microbiol Infect Dis 9:717–724

    Article  PubMed  CAS  Google Scholar 

  16. Huletsky A, Giroux R, Rossbach V, Gagnon M, Vaillancourt M, Bernier M, Gagnon F, Truchon K, Bastien M, Picard FJ, van BA, Ouellette M, Roy PH, Bergeron MG (2004) New real-time PCR assay for rapid detection of methicillin-resistant Staphylococcus aureus directly from specimens containing a mixture of staphylococci. J Clin Microbiol 42:1875–1884

    Article  PubMed  CAS  Google Scholar 

  17. Hartmann H, Stender H, Schäfer A, Autenrieth IB, Kempf VA (2005) Rapid identification of Staphylococcus aureus in blood cultures by a combination of fluorescence in situ hybridization using peptide nucleic acid probes and flow cytometry. J Clin Microbiol 43:4855–4857

    Article  PubMed  CAS  Google Scholar 

  18. Thomas LC, Gidding HF, Ginn AN, Olma T, Iredell J (2007) Development of a real-time Staphylococcus aureus and MRSA (SAM-) PCR for routine blood culture. J Microbiol Methods 68:296–302

    Article  PubMed  CAS  Google Scholar 

  19. Kempf VA, Trebesius K, Autenrieth IB (2000) Fluorescent In situ hybridization allows rapid identification of microorganisms in blood cultures. J Clin Microbiol 38:830–838

    PubMed  CAS  Google Scholar 

Download references

Acknowledgement

The authors thank Josef Albus, Ingrid Cheru, Cornelia Lüth and Stefanie Richt for excellent technical assistance.

Author information

Authors and Affiliations

  1. Institut für Medizinische Mikrobiologie und Hygiene, Elfriede Aulhorn Str. 6, 72076, Tübingen, Germany

    S. Gröbner & V. A. J. Kempf

Authors
  1. S. Gröbner
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. V. A. J. Kempf
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to V. A. J. Kempf.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Gröbner, S., Kempf, V.A.J. Rapid detection of methicillin-resistant staphylococci by real-time PCR directly from positive blood culture bottles. Eur J Clin Microbiol Infect Dis 26, 751–754 (2007). https://doi.org/10.1007/s10096-007-0368-3

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10096-007-0368-3

Keywords

Search

Navigation