Skip to content
1887

Journal of Medical Microbiology logo Journal of Medical Microbiology

Volume 66, Issue 12

Short-term incubation of positive blood cultures in brain-heart infusion broth accelerates identification of bacteria by matrix-assisted laser desorption/ionization time-of-flight mass-spectrometry No Access

Abstract

Fast identification of bacteria directly from positive blood cultures (BCs) by matrix-assisted laser desorption/ionization time-of-flight mass-spectrometry (MALDI-TOF MS) can be achieved either using the MALDI Sepsityper kit (protein extraction method) or after a short-term pre-cultivation step on solid medium. We developed a new method that involves short-term enrichment of positive BCs in brain–heart infusion broth (BHI) prior to MALDI-TOF MS analysis.

Eighty-four BCs flagged as positive were included in this study; these were processed in parallel either directly using the MALDI Sepsityper kit or following a short-term culture either in BHI or on Columbia blood agar with 5 % sheep blood (CBA).

Bacterial species were successfully identified in 91.6, 89.2 and 65.4 % of cases after pre-cultivation for 4 h in BHI, on CBA, or by using the MALDI Sepsityper kit, respectively. Overall, the mean incubation time to correct identification was shorter when pre-cultures were performed in BHI; the mean time for Gram-negative rods was 78.2 min in BHI and 108.2 min on CBA (=0.045), and the mean time for Gram-positive cocci was 128.5 min in BHI and 169.6 min on CBA (=0.013).

Short-term enrichment of BCs in BHI accelerates identification of a number of bacterial species by MALDI-TOF MS. Further prospective studies are needed to validate our method and gauge its potential clinical impact on the management of bloodstream bacterial infections.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): blood cultures , broth incubation , fast identification , MALDI-TOF MS and short-term incubation
© 2017 The Authors
Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.000643
2017-12-01
2025-02-10
Loading full text...

Full text loading...

References

  1. Ibrahim EH, Sherman G, Ward S, Fraser VJ, Kollef MH. The influence of inadequate antimicrobial treatment of bloodstream infections on patient outcomes in the ICU setting. Chest 2000; 118:146–155 [View Article][PubMed]
     [Google Scholar]
  2. Kumar A, Roberts D, Wood KE, Light B, Parrillo JE et al. Duration of hypotension before initiation of effective antimicrobial therapy is the critical determinant of survival in human septic shock. Crit Care Med 2006; 34:1589–1596 [View Article][PubMed]
     [Google Scholar]
  3. Fraser A, Paul M, Almanasreh N, Tacconelli E, Frank U et al. Benefit of appropriate empirical antibiotic treatment: thirty-day mortality and duration of hospital stay. Am J Med 2006; 119:970–976 [View Article][PubMed]
     [Google Scholar]
  4. La Scola B, Raoult D. Direct identification of bacteria in positive blood culture bottles by matrix-assisted laser desorption ionisation time-of-flight mass spectrometry. PLoS One 2009; 4:e8041 [View Article][PubMed]
     [Google Scholar]
  5. Stevenson LG, Drake SK, Murray PR. Rapid identification of bacteria in positive blood culture broths by matrix-assisted laser desorption ionization-time of flight mass spectrometry. J Clin Microbiol 2010; 48:444–447 [View Article][PubMed]
     [Google Scholar]
  6. Klein S, Zimmermann S, Köhler C, Mischnik A, Alle W et al. Integration of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry in blood culture diagnostics: a fast and effective approach. J Med Microbiol 2012; 61:323–331 [View Article][PubMed]
     [Google Scholar]
  7. Lagacé-Wiens PR, Adam HJ, Karlowsky JA, Nichol KA, Pang PF et al. Identification of blood culture isolates directly from positive blood cultures by use of matrix-assisted laser desorption ionization-time of flight mass spectrometry and a commercial extraction system: analysis of performance, cost, and turnaround time. J Clin Microbiol 2012; 50:3324–3328 [View Article][PubMed]
     [Google Scholar]
  8. Idelevich EA, Schüle I, Grünastel B, Wüllenweber J, Peters G et al. Rapid identification of microorganisms from positive blood cultures by MALDI-TOF mass spectrometry subsequent to very short-term incubation on solid medium. Clin Microbiol Infect 2014; 20:1001–1006 [View Article][PubMed]
     [Google Scholar]
  9. Kohlmann R, Hoffmann A, Geis G, Gatermann S. MALDI-TOF mass spectrometry following short incubation on a solid medium is a valuable tool for rapid pathogen identification from positive blood cultures. Int J Med Microbiol 2015; 305:469–479 [View Article][PubMed]
     [Google Scholar]
  10. Zabbe JB, Zanardo L, Mégraud F, Bessède E. MALDI-TOF mass spectrometry for early identification of bacteria grown in blood culture bottles. J Microbiol Methods 2015; 115:45–46 [View Article][PubMed]
     [Google Scholar]
  11. Verroken A, Defourny L, Lechgar L, Magnette A, Delmée M et al. Reducing time to identification of positive blood cultures with MALDI-TOF MS analysis after a 5-h subculture. Eur J Clin Microbiol Infect Dis 2015; 34:405–413 [View Article][PubMed]
     [Google Scholar]
  12. Chen Y, Porter V, Mubareka S, Kotowich L, Simor AE. Rapid identification of bacteria directly from positive blood cultures by use of a serum separator tube, smudge plate preparation, and matrix-assisted laser desorption ionization-time of flight mass spectrometry. J Clin Microbiol 2015; 53:3349–3352 [View Article][PubMed]
     [Google Scholar]
  13. Bhatti MM, Boonlayangoor S, Beavis KG, Tesic V. Rapid identification of positive blood cultures by matrix-assisted laser desorption ionization-time of flight mass spectrometry using prewarmed agar plates. J Clin Microbiol 2014; 52:4334–4338 [View Article][PubMed]
     [Google Scholar]
  14. Curtoni A, Cipriani R, Marra ES, Barbui AM, Cavallo R et al. Rapid identification of microorganisms from positive blood culture by MALDI-TOF MS after short-term incubation on solid medium. Curr Microbiol 2017; 74:97–102 [View Article][PubMed]
     [Google Scholar]
  15. Delport JA, Strikwerda A, Armstrong A, Schaus D, John M. Quality of care is improved by rapid short incubation MALDI-ToF identification from blood cultures as measured by reduced length of stay and patient outcomes as part of a multi-disciplinary approach to bacteremia in pediatric patients. PLoS One 2016; 11:e0160618 [View Article][PubMed]
     [Google Scholar]
  16. Delport JA, Strikwerda A, Armstrong A, Schaus D, John M. MALDI-ToF short incubation identification from blood cultures is associated with reduced length of hospitalization and a decrease in bacteremia associated mortality. Eur J Clin Microbiol Infect Dis 2017; 36:1181–1186 [View Article][PubMed]
     [Google Scholar]
  17. Kroumova V, Gobbato E, Basso E, Mucedola L, Giani T et al. Direct identification of bacteria in blood culture by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry: a new methodological approach. Rapid Commun Mass Spectrom 2011; 25:2247–2249 [View Article][PubMed]
     [Google Scholar]
  18. Buchan BW, Riebe KM, Ledeboer NA. Comparison of the MALDI Biotyper system using Sepsityper specimen processing to routine microbiological methods for identification of bacteria from positive blood culture bottles. J Clin Microbiol 2012; 50:346–352 [View Article][PubMed]
     [Google Scholar]
  19. Wieser A, Schneider L, Jung J, Schubert S. MALDI-TOF MS in microbiological diagnostics-identification of microorganisms and beyond (mini review). Appl Microbiol Biotechnol 2012; 93:965–974 [View Article][PubMed]
     [Google Scholar]
  20. Buehler SS, Madison B, Snyder SR, Derzon JH, Cornish NE et al. Effectiveness of practices to increase timeliness of providing targeted therapy for inpatients with bloodstream infections: a laboratory medicine best practices systematic review and meta-analysis. Clin Microbiol Rev 2016; 29:59–103 [View Article][PubMed]
     [Google Scholar]
  21. Kok J, Thomas LC, Olma T, Chen SC, Iredell JR. Identification of bacteria in blood culture broths using matrix-assisted laser desorption-ionization Sepsityper™ and time of flight mass spectrometry. PLoS One 2011; 6:e23285 [View Article][PubMed]
     [Google Scholar]
  22. Matsuda N, Matsuda M, Notake S, Yokokawa H, Kawamura Y et al. Evaluation of a simple protein extraction method for species identification of clinically relevant staphylococci by matrix-assisted laser desorption ionization-time of flight mass spectrometry. J Clin Microbiol 2012; 50:3862–3866 [View Article][PubMed]
     [Google Scholar]
  23. Martiny D, Dediste A, Vandenberg O. Comparison of an in-house method and the commercial Sepsityper™ kit for bacterial identification directly from positive blood culture broths by matrix-assisted laser desorption-ionisation time-of-flight mass spectrometry. Eur J Clin Microbiol Infect Dis 2012; 31:2269–2281 [View Article][PubMed]
     [Google Scholar]
  24. Maelegheer K, Nulens E. Same-day identification and antibiotic susceptibility testing on positive blood cultures: a simple and inexpensive procedure. Eur J Clin Microbiol Infect Dis 2017; 36:681–687 [View Article][PubMed]
     [Google Scholar]
  25. Clerc O, Prod'hom G, Vogne C, Bizzini A, Calandra T et al. Impact of matrix-assisted laser desorption ionization time-of-flight mass spectrometry on the clinical management of patients with Gram-negative bacteremia: a prospective observational study. Clin Infect Dis 2013; 56:1101–1107 [View Article][PubMed]
     [Google Scholar]
/content/journal/jmm/10.1099/jmm.0.000643
Loading
Short-term incubation of positive blood cultures in brain-heart infusion broth accelerates identification of bacteria by matrix-assisted laser desorption/ionization time-of-flight mass-spectrometry
J. Med. Microbiol. 66, 1752 (2017); https://doi.org/10.1099/jmm.0.000643
/content/journal/jmm/10.1099/jmm.0.000643
/content/journal/jmm/10.1099/jmm.0.000643
Loading

Data & Media loading...

Most cited Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error