Skip to main content
Top
Published in: European Journal of Clinical Microbiology & Infectious Diseases 2/2015

01-02-2015 | Article

Reducing time to identification of positive blood cultures with MALDI-TOF MS analysis after a 5-h subculture

Authors: A. Verroken, L. Defourny, L. Lechgar, A. Magnette, M. Delmée, Y. Glupczynski

Published in: European Journal of Clinical Microbiology & Infectious Diseases | Issue 2/2015

Login to get access

Abstract

Speeding up the turn-around time of positive blood culture identifications is essential in order to optimize the treatment of septic patients. Several sample preparation techniques have been developed allowing direct matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) identification of positive blood cultures. Yet, the hands-on time restrains their routine workflow. In this study, we evaluated an approach whereby MALDI-TOF MS identification without any additional steps was carried out on short subcultured colonies from positive blood bottles with the objective of allowing results reporting on the day of positivity detection. Over a 7-month period in 2012, positive blood cultures detected by 9 am with an automated system were inoculated onto a Columbia blood agar and processed after a 5-h incubation on a MALDI-TOF MicroFlex platform (Bruker Daltonik GmbH). Single-spotted colonies were covered with 1 μl formic acid and 1 μl matrix solution. The results were compared to the validated identification techniques. A total of 925 positive blood culture bottles (representing 470 bacteremic episodes) were included. Concordant identification was obtained in 727 (81.1 %) of the 896 monomicrobial blood cultures, with failure being mostly observed with anaerobes and yeasts. In 17 episodes of polymicrobic bacteremia, the identification of one of the two isolates was achieved in 24/29 (82.7 %) positive cultures. Routine implementation of MALDI-TOF MS identification on young positive blood subcultures provides correct results to the clinician in more than 80 % of the bacteremic episodes and allows access to identification results on the day of blood culture positivity detection, potentially accelerating the implementation of targeted clinical treatments.
Literature
1.
go back to reference 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 41:3119–3125CrossRefPubMedCentralPubMed 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 41:3119–3125CrossRefPubMedCentralPubMed
2.
go back to reference Galar A, Leiva J, Espinosa M, Guillén-Grima F, Hernáez S, Yuste JR (2012) Clinical and economic evaluation of the impact of rapid microbiological diagnostic testing. J Infect 65:302–309CrossRefPubMed Galar A, Leiva J, Espinosa M, Guillén-Grima F, Hernáez S, Yuste JR (2012) Clinical and economic evaluation of the impact of rapid microbiological diagnostic testing. J Infect 65:302–309CrossRefPubMed
3.
go back to reference Stoneking LR, Patanwala AE, Winkler JP, Fiorello AB, Lee ES, Olson DP, Wolk DM (2013) Would earlier microbe identification alter antibiotic therapy in bacteremic emergency department patients? J Emerg Med 44:1–8CrossRefPubMed Stoneking LR, Patanwala AE, Winkler JP, Fiorello AB, Lee ES, Olson DP, Wolk DM (2013) Would earlier microbe identification alter antibiotic therapy in bacteremic emergency department patients? J Emerg Med 44:1–8CrossRefPubMed
4.
go back to reference Peters RPH, van Agtmael MA, Danner SA, Savelkoul PHM, Vandenbroucke-Grauls CMJE (2004) New developments in the diagnosis of bloodstream infections. Lancet Infect Dis 4:751–760CrossRefPubMed Peters RPH, van Agtmael MA, Danner SA, Savelkoul PHM, Vandenbroucke-Grauls CMJE (2004) New developments in the diagnosis of bloodstream infections. Lancet Infect Dis 4:751–760CrossRefPubMed
6.
go back to reference Seng P, Drancourt M, Gouriet F, La Scola B, Fournier PE, Rolain JM, Raoult D (2009) Ongoing revolution in bacteriology: routine identification of bacteria by matrix-assisted laser desorption ionization time-of-flight mass spectrometry. Clin Infect Dis 49:543–551CrossRefPubMed Seng P, Drancourt M, Gouriet F, La Scola B, Fournier PE, Rolain JM, Raoult D (2009) Ongoing revolution in bacteriology: routine identification of bacteria by matrix-assisted laser desorption ionization time-of-flight mass spectrometry. Clin Infect Dis 49:543–551CrossRefPubMed
7.
go back to reference Ferroni A, Suarez S, Beretti JL, Dauphin B, Bille E, Meyer J, Bougnoux ME, Alanio A, Berche P, Nassif X (2010) Real-time identification of bacteria and Candida species in positive blood culture broths by matrix-assisted laser desorption ionization-time of flight mass spectrometry. J Clin Microbiol 48:1542–1548CrossRefPubMedCentralPubMed Ferroni A, Suarez S, Beretti JL, Dauphin B, Bille E, Meyer J, Bougnoux ME, Alanio A, Berche P, Nassif X (2010) Real-time identification of bacteria and Candida species in positive blood culture broths by matrix-assisted laser desorption ionization-time of flight mass spectrometry. J Clin Microbiol 48:1542–1548CrossRefPubMedCentralPubMed
8.
go back to reference Fuglsang-Damgaard D, Houlberg Nielsen C, Mandrup E, Fuursted K (2011) The use of Gram stain and matrix-assisted laser desorption ionization time-of-flight mass spectrometry on positive blood culture: synergy between new and old technology. APMIS 119:681–688CrossRefPubMed Fuglsang-Damgaard D, Houlberg Nielsen C, Mandrup E, Fuursted K (2011) The use of Gram stain and matrix-assisted laser desorption ionization time-of-flight mass spectrometry on positive blood culture: synergy between new and old technology. APMIS 119:681–688CrossRefPubMed
9.
go back to reference Loonen AJM, Jansz AR, Stalpers J, Wolffs PFG, van den Brule AJC (2012) An evaluation of three processing methods and the effect of reduced culture times for faster direct identification of pathogens from BacT/ALERT blood cultures by MALDI-TOF MS. Eur J Clin Microbiol Infect Dis 31:1575–1583CrossRefPubMedCentralPubMed Loonen AJM, Jansz AR, Stalpers J, Wolffs PFG, van den Brule AJC (2012) An evaluation of three processing methods and the effect of reduced culture times for faster direct identification of pathogens from BacT/ALERT blood cultures by MALDI-TOF MS. Eur J Clin Microbiol Infect Dis 31:1575–1583CrossRefPubMedCentralPubMed
10.
go back to reference Martiny D, Dediste A, Vandenberg O (2012) 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 31:2269–2281CrossRefPubMed Martiny D, Dediste A, Vandenberg O (2012) 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 31:2269–2281CrossRefPubMed
11.
go back to reference Buchan BW, Riebe KM, Ledeboer NA (2012) 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 50:346–352CrossRefPubMedCentralPubMed Buchan BW, Riebe KM, Ledeboer NA (2012) 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 50:346–352CrossRefPubMedCentralPubMed
12.
go back to reference Meex C, Neuville F, Descy J, Huynen P, Hayette MP, De Mol P, Melin P (2012) Direct identification of bacteria from BacT/ALERT anaerobic positive blood cultures by MALDI-TOF MS: MALDI Sepsityper kit versus an in-house saponin method for bacterial extraction. J Med Microbiol 61:1511–1516CrossRefPubMed Meex C, Neuville F, Descy J, Huynen P, Hayette MP, De Mol P, Melin P (2012) Direct identification of bacteria from BacT/ALERT anaerobic positive blood cultures by MALDI-TOF MS: MALDI Sepsityper kit versus an in-house saponin method for bacterial extraction. J Med Microbiol 61:1511–1516CrossRefPubMed
13.
go back to reference Romero-Gómez MP, Gómez-Gil R, Paño-Pardo JR, Mingorance J (2012) Identification and susceptibility testing of microorganism by direct inoculation from positive blood culture bottles by combining MALDI-TOF and Vitek-2 Compact is rapid and effective. J Infect 65:513–520CrossRefPubMed Romero-Gómez MP, Gómez-Gil R, Paño-Pardo JR, Mingorance J (2012) Identification and susceptibility testing of microorganism by direct inoculation from positive blood culture bottles by combining MALDI-TOF and Vitek-2 Compact is rapid and effective. J Infect 65:513–520CrossRefPubMed
14.
go back to reference Chen JHK, Ho PL, Kwan GSW, She KKK, Siu GKH, Cheng VCC, Yuen KY, Yam WC (2013) Direct bacterial identification in positive blood cultures by use of two commercial matrix-assisted laser desorption ionization-time of flight mass spectrometry systems. J Clin Microbiol 51:1733–1739CrossRefPubMedCentralPubMed Chen JHK, Ho PL, Kwan GSW, She KKK, Siu GKH, Cheng VCC, Yuen KY, Yam WC (2013) Direct bacterial identification in positive blood cultures by use of two commercial matrix-assisted laser desorption ionization-time of flight mass spectrometry systems. J Clin Microbiol 51:1733–1739CrossRefPubMedCentralPubMed
15.
go back to reference Leli C, Cenci E, Cardaccia A, Moretti A, D’Alò F, Pagliochini R, Barcaccia M, Farinelli S, Vento S, Bistoni F, Mencacci A (2013) Rapid identification of bacterial and fungal pathogens from positive blood cultures by MALDI-TOF MS. Int J Med Microbiol 303:205–209CrossRefPubMed Leli C, Cenci E, Cardaccia A, Moretti A, D’Alò F, Pagliochini R, Barcaccia M, Farinelli S, Vento S, Bistoni F, Mencacci A (2013) Rapid identification of bacterial and fungal pathogens from positive blood cultures by MALDI-TOF MS. Int J Med Microbiol 303:205–209CrossRefPubMed
16.
go back to reference Martiny D, Debaugnies F, Gateff D, Gérard M, Aoun M, Martin C, Konopnicki D, Loizidou A, Georgala A, Hainaut M, Chantrenne M, Dediste A, Vandenberg O, Van Praet S (2013) Impact of rapid microbial identification directly from positive blood cultures using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry on patient management. Clin Microbiol Infect 19:E568–E581CrossRefPubMed Martiny D, Debaugnies F, Gateff D, Gérard M, Aoun M, Martin C, Konopnicki D, Loizidou A, Georgala A, Hainaut M, Chantrenne M, Dediste A, Vandenberg O, Van Praet S (2013) Impact of rapid microbial identification directly from positive blood cultures using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry on patient management. Clin Microbiol Infect 19:E568–E581CrossRefPubMed
18.
go back to reference De Bel A, Wybo I, Piérard D, Lauwers S (2010) Correct implementation of matrix-assisted laser desorption ionization-time of flight mass spectrometry in routine clinical microbiology. J Clin Microbiol 48:1991–1992CrossRefPubMedCentralPubMed De Bel A, Wybo I, Piérard D, Lauwers S (2010) Correct implementation of matrix-assisted laser desorption ionization-time of flight mass spectrometry in routine clinical microbiology. J Clin Microbiol 48:1991–1992CrossRefPubMedCentralPubMed
19.
go back to reference Wauters G, Avesani V, Laffineur K, Charlier J, Janssens M, Van Bosterhaut B, Delmée M (2003) Brevibacterium lutescens sp. nov., from human and environmental samples. Int J Syst Evol Microbiol 53:1321–1325CrossRefPubMed Wauters G, Avesani V, Laffineur K, Charlier J, Janssens M, Van Bosterhaut B, Delmée M (2003) Brevibacterium lutescens sp. nov., from human and environmental samples. Int J Syst Evol Microbiol 53:1321–1325CrossRefPubMed
21.
go back to reference McElvania TeKippe E, Shuey S, Winkler DW, Butler MA, Burnham CAD (2013) Optimizing identification of clinically relevant Gram-positive organisms by use of the Bruker Biotyper matrix-assisted laser desorption ionization-time of flight mass spectrometry system. J Clin Microbiol 51:1421–1427CrossRefPubMedCentralPubMed McElvania TeKippe E, Shuey S, Winkler DW, Butler MA, Burnham CAD (2013) Optimizing identification of clinically relevant Gram-positive organisms by use of the Bruker Biotyper matrix-assisted laser desorption ionization-time of flight mass spectrometry system. J Clin Microbiol 51:1421–1427CrossRefPubMedCentralPubMed
22.
go back to reference Ford BA, Burnham CAD (2013) Optimization of routine identification of clinically relevant Gram-negative bacteria by use of matrix-assisted laser desorption ionization-time of flight mass spectrometry and the Bruker Biotyper. J Clin Microbiol 51:1412–1420CrossRefPubMedCentralPubMed Ford BA, Burnham CAD (2013) Optimization of routine identification of clinically relevant Gram-negative bacteria by use of matrix-assisted laser desorption ionization-time of flight mass spectrometry and the Bruker Biotyper. J Clin Microbiol 51:1412–1420CrossRefPubMedCentralPubMed
23.
go back to reference Idelevich EA, Schüle I, Grünastel B, Wüllenweber J, Peters G, Becker K (2014) 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. doi:10.1111/1469-0691.12640 Idelevich EA, Schüle I, Grünastel B, Wüllenweber J, Peters G, Becker K (2014) 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. doi:10.​1111/​1469-0691.​12640
24.
go back to reference Mestas J, Felsenstein S, Dien Bard J (2014) Direct identification of bacteria from positive BacT/ALERT blood culture bottles using matrix-assisted laser desorption ionization-time-of-flight mass spectrometry. Diagn Microbiol Infect Dis. doi:10.1016/j.diagmicrobio.2014.07.008 PubMed Mestas J, Felsenstein S, Dien Bard J (2014) Direct identification of bacteria from positive BacT/ALERT blood culture bottles using matrix-assisted laser desorption ionization-time-of-flight mass spectrometry. Diagn Microbiol Infect Dis. doi:10.​1016/​j.​diagmicrobio.​2014.​07.​008 PubMed
25.
go back to reference Bassetti M, Merelli M, Righi E, Diaz-Martin A, Rosello EM, Luzzati R, Parra A, Trecarichi EM, Sanguinetti M, Posteraro B, Garnacho-Montero J, Sartor A, Rello J, Tumbarello M (2013) Epidemiology, species distribution, antifungal susceptibility, and outcome of candidemia across five sites in Italy and Spain. J Clin Microbiol 51:4167–4172CrossRefPubMedCentralPubMed Bassetti M, Merelli M, Righi E, Diaz-Martin A, Rosello EM, Luzzati R, Parra A, Trecarichi EM, Sanguinetti M, Posteraro B, Garnacho-Montero J, Sartor A, Rello J, Tumbarello M (2013) Epidemiology, species distribution, antifungal susceptibility, and outcome of candidemia across five sites in Italy and Spain. J Clin Microbiol 51:4167–4172CrossRefPubMedCentralPubMed
26.
go back to reference Ngo JT, Parkins MD, Gregson DB, Pitout JD, Ross T, Church DL, Laupland KB (2013) Population-based assessment of the incidence, risk factors, and outcomes of anaerobic bloodstream infections. Infection 41:41–48CrossRefPubMed Ngo JT, Parkins MD, Gregson DB, Pitout JD, Ross T, Church DL, Laupland KB (2013) Population-based assessment of the incidence, risk factors, and outcomes of anaerobic bloodstream infections. Infection 41:41–48CrossRefPubMed
27.
go back to reference Pulcrano G, Iula DV, Vollaro A, Tucci A, Cerullo M, Esposito M, Rossano F, Catania MR (2013) Rapid and reliable MALDI-TOF mass spectrometry identification of Candida non-albicans isolates from bloodstream infections. J Microbiol Methods 94:262–266CrossRefPubMed Pulcrano G, Iula DV, Vollaro A, Tucci A, Cerullo M, Esposito M, Rossano F, Catania MR (2013) Rapid and reliable MALDI-TOF mass spectrometry identification of Candida non-albicans isolates from bloodstream infections. J Microbiol Methods 94:262–266CrossRefPubMed
28.
go back to reference Neville SA, LeCordier A, Ziochos H, Chater MJ, Gosbell IB, Maley MW, van Hal SJ (2011) Utility of matrix-assisted laser desorption ionization-time of flight mass spectrometry following introduction for routine laboratory bacterial identification. J Clin Microbiol 49:2980–2984CrossRefPubMedCentralPubMed Neville SA, LeCordier A, Ziochos H, Chater MJ, Gosbell IB, Maley MW, van Hal SJ (2011) Utility of matrix-assisted laser desorption ionization-time of flight mass spectrometry following introduction for routine laboratory bacterial identification. J Clin Microbiol 49:2980–2984CrossRefPubMedCentralPubMed
29.
go back to reference Kärpänoja P, Harju I, Rantakokko-Jalava K, Haanperä M, Sarkkinen H (2014) Evaluation of two matrix-assisted laser desorption ionization-time of flight mass spectrometry systems for identification of viridans group streptococci. Eur J Clin Microbiol Infect Dis 33:779–788. doi:10.1007/s10096-013-2012-8 CrossRefPubMed Kärpänoja P, Harju I, Rantakokko-Jalava K, Haanperä M, Sarkkinen H (2014) Evaluation of two matrix-assisted laser desorption ionization-time of flight mass spectrometry systems for identification of viridans group streptococci. Eur J Clin Microbiol Infect Dis 33:779–788. doi:10.​1007/​s10096-013-2012-8 CrossRefPubMed
Metadata
Title
Reducing time to identification of positive blood cultures with MALDI-TOF MS analysis after a 5-h subculture
Authors
A. Verroken
L. Defourny
L. Lechgar
A. Magnette
M. Delmée
Y. Glupczynski
Publication date
01-02-2015
Publisher
Springer Berlin Heidelberg
Published in
European Journal of Clinical Microbiology & Infectious Diseases / Issue 2/2015
Print ISSN: 0934-9723
Electronic ISSN: 1435-4373
DOI
https://doi.org/10.1007/s10096-014-2242-4

Other articles of this Issue 2/2015

European Journal of Clinical Microbiology & Infectious Diseases 2/2015 Go to the issue
Live Webinar | 27-06-2024 | 18:00 (CEST)

Keynote webinar | Spotlight on medication adherence

Live: Thursday 27th June 2024, 18:00-19:30 (CEST)

WHO estimates that half of all patients worldwide are non-adherent to their prescribed medication. The consequences of poor adherence can be catastrophic, on both the individual and population level.

Join our expert panel to discover why you need to understand the drivers of non-adherence in your patients, and how you can optimize medication adherence in your clinics to drastically improve patient outcomes.

Prof. Kevin Dolgin
Prof. Florian Limbourg
Prof. Anoop Chauhan
Developed by: Springer Medicine
Obesity Clinical Trial Summary

At a glance: The STEP trials

A round-up of the STEP phase 3 clinical trials evaluating semaglutide for weight loss in people with overweight or obesity.

Developed by: Springer Medicine

Highlights from the ACC 2024 Congress

Year in Review: Pediatric cardiology

Watch Dr. Anne Marie Valente present the last year's highlights in pediatric and congenital heart disease in the official ACC.24 Year in Review session.

Year in Review: Pulmonary vascular disease

The last year's highlights in pulmonary vascular disease are presented by Dr. Jane Leopold in this official video from ACC.24.

Year in Review: Valvular heart disease

Watch Prof. William Zoghbi present the last year's highlights in valvular heart disease from the official ACC.24 Year in Review session.

Year in Review: Heart failure and cardiomyopathies

Watch this official video from ACC.24. Dr. Biykem Bozkurt discusses last year's major advances in heart failure and cardiomyopathies.