1887

Abstract

Rapid identification of the causative agent of sepsis is crucial for patient outcomes.

. The Sepsityper sample preparation method enables direct microbial identification of positive blood culture samples via matrix-assisted laser desorption ionization/time-of-flight mass spectrometry (MALDI-TOF MS).

. The implementation of the Sepsityper method in the routine practice could represent a fundamental tool to achieve a prompt identification of the causative agent of bloodstream infections, and therefore accelerate the adoption of the proper antibiotic treatment.

In this study, the novel rapid workflow of the MALDI Biotypr Sepsityper kit (Bruker Daltonik GmbH, Germany) was evaluated using routine samples from a 2-year period (=6918), and dedicated optimized protocols for the microbial groups that were more difficult to identify were developed. Moreover, the use of the residual bacterial pellet to perform susceptibility testing using different methods (commercial broth microdilution, disc diffusion, gradient diffusion) was investigated.

The rapid Sepsityper protocol allowed the identification of 5470/6338 (86.3 %) monomicrobial samples at species level, with very good performance for all of the clinically most significant pathogens (2510/2592 enterobacteria, 631/669 and 223/246 enterococci were identified). , and yeasts were the most troublesome to identify, but the application of specific optimized protocols significantly improved their rate of identification (from 14.7–71.5 %, 47.8–89.7 % and 37.1–89.5 %, respectively). Specificity was 100 % (no identification was made for the false-positive samples). Further, the residual pellet proved to be suitable to investigate susceptibility to antimicrobials, enabling us to simplify the workflow and shorten the time to report.

The Rapid Sepsityper workflow proved to be a reliable sample preparation method for identification and susceptibility testing directly from positive blood cultures, providing novel approaches for accelerated diagnostics of bloodstream infections.

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2020-11-06
2024-04-19
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References

  1. Adhikari NKJ, Fowler RA, Bhagwanjee S, Rubenfeld GD. Critical care and the global burden of critical illness in adults. The Lancet 2010; 376:1339–1346 [View Article]
    [Google Scholar]
  2. Rhodes A, Evans LE, Alhazzani W, Levy MM, Antonelli M et al. Surviving sepsis campaign: international guidelines for management of sepsis and septic shock: 2016. Intensive Care Med 2017; 43:304–377 [View Article]
    [Google Scholar]
  3. 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]
    [Google Scholar]
  4. Chaubey VP, Pitout JD, Dalton B, Ross T, Church DL et al. Clinical outcome of empiric antimicrobial therapy of bacteremia due to extended-spectrum beta-lactamase producing Escherichia coli and Klebsiella pneumoniae. BMC Res Notes 2010; 27:116
    [Google Scholar]
  5. Paul M, Shani V, Muchtar E, Kariv G, Robenshtok E et al. Systematic review and meta-analysis of the efficacy of appropriate empiric antibiotic therapy for sepsis. Antimicrob Agents Chemother 2010; 54:4851–4863 [View Article]
    [Google Scholar]
  6. Taur Y, Cohen N, Dubnow S, Paskovaty A, Seo SK. Effect of antifungal therapy timing on mortality in cancer patients with candidemia. Antimicrob Agents Chemother 2010; 54:184–190 [View Article]
    [Google Scholar]
  7. Pogue JM, Kaye KS, Cohen DA, Marchaim D. Appropriate antimicrobial therapy in the era of multidrug-resistant human pathogens. Clin Microbiol Infect 2015; 21:302–312 [View Article]
    [Google Scholar]
  8. Lagace-Wiens PRS, 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]
    [Google Scholar]
  9. Faron ML, Buchan BW, Ledeboer NA. Matrix-Assisted laser desorption Ionization–Time of flight mass spectrometry for use with positive blood cultures: methodology, performance, and optimization. J Clin Microbiol 2017; 55:3328–3338 [View Article]
    [Google Scholar]
  10. Schubert S, Weinert K, Wagner C, Gunzl B, Wieser A et al. Novel, improved sample preparation for rapid, direct identification from positive blood cultures using matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry. J Mol Diagn 2011; 13:701–706 [View Article]
    [Google Scholar]
  11. Szabados F, Michels M, Kaase M, Gatermann S. The sensitivity of direct identification from positive BacT/ALERT (bioMérieux) blood culture bottles by matrix-assisted laser desorption ionization time-of-flight mass spectrometry is low. Clin Microb and Infection 2011; 17:192–195 [View Article]
    [Google Scholar]
  12. Schmidt V, Jarosch A, März P, Sander C, Vacata V et al. Rapid identification of bacteria in positive blood culture by matrix-assisted laser desorption ionization time-of-flight mass spectrometry. Eur J Clin Microb and Infect Dis 2012; 31:311–317 [View Article]
    [Google Scholar]
  13. Haigh JD, Green IM, Ball D, Eydmann M, Millar M et al. Rapid identification of bacteria from bioMerieux BacT/Alert blood culture bottles by MALDI-TOF MS. Br J Biomed Sci 2013; 70:149–155 [View Article]
    [Google Scholar]
  14. Morgenthaler NG, Kostrzewa M. Rapid identification of pathogens in positive blood culture of patients with sepsis: review and meta-analysis of the performance of the Sepsityper kit. Int J Microbiol 2015; 2015:827416 [View Article]
    [Google Scholar]
  15. Wimmer JL, Long SW, Cernoch P, Land GA, Davis JR et al. Strategy for rapid identification and antibiotic susceptibility testing of gram-negative bacteria directly recovered from positive blood cultures using the Bruker MALDI Biotyper and the BD Phoenix system. J Clin Microbiol 2012; 50:2452–2454 [View Article]
    [Google Scholar]
  16. Hazelton B, Thomas LC, Olma T, Kok J, O’Sullivan M et al. Rapid and accurate direct antibiotic susceptibility testing of blood culture broths using MALDI Sepsityper combined with the BD Phoenix automated system. J Med Microbiol 2014; 63:1590–1594 [View Article]
    [Google Scholar]
  17. Idelevich EA, Grunewald CM, Wüllenweber J, Becker K. Rapid identification and susceptibility testing of Candida spp. from positive blood cultures by combination of direct MALDI-TOF mass spectrometry and direct inoculation of Vitek 2. PLoS One 2014; 9:e114834 [View Article]
    [Google Scholar]
  18. Christner M, Rohde H, Wolters M, Sobottka I, Wegscheider K et al. Rapid identification of bacteria from positive blood culture bottles by use of matrix-assisted laser desorption-ionization time of flight mass spectrometry fingerprinting. J Clin Microbiol 2010; 48:1584–1591 [View Article]
    [Google Scholar]
  19. Scohy A, Noël A, Boeras A, Brassinne L, Laurent T et al. Evaluation of the Bruker MBT Sepsityper IVD module for the identification of polymicrobial blood cultures with MALDI-TOF MS. Eur J Clin Microbiol Infect Dis 2018; 37:2145–2152 [View Article]
    [Google Scholar]
  20. Cordovana M, Kostrzewa M, Glandorf J, Bienia M, Ambretti S et al. A full MALDI-based approach to detect plasmid-encoded KPC-producing Klebsiella pneumoniae. Front Microbiol 2018; 23:2854
    [Google Scholar]
  21. Hu Y, Huang Y, Lizou Y, Li J, Zhang R. Evaluation of Staphylococcus aureus subtyping module for methicillin-resistant Staphylococcus aureus detection based on matrix-assisted laser desorption ionization time-of-flight mass spectrometry. Front Microbiol 2019; 31:2504
    [Google Scholar]
  22. Clinical and Laboratory Standards Institute (CLSI) Laboratory detection and identification of Mycobacteria. Clsi Guideline M48, 2nd ed. Wayne, PA: Clinical and Laboratory Standards Institute; 2018
    [Google Scholar]
  23. Munson EL, Diekema DJ, Beekmann SE, Chapin KC, Doern GV. Detection and treatment of bloodstream infection: laboratory reporting and antimicrobial management. J Clin Microbiol 2003; 41:495–497 [View Article]
    [Google Scholar]
  24. Hazelton B, Thomas LC, Olma T, Kok J, O’Sullivan M et al. Rapid and accurate direct antibiotic susceptibility testing of blood culture broths using MALDI Sepsityper combined with the BD Phoenix automated system. J Med Microbiol 2014; 63:1590–1594 [View Article]
    [Google Scholar]
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