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Published in: European Journal of Clinical Microbiology & Infectious Diseases 6/2020

01-06-2020 | Original Article

Rapid identification by MALDI-TOF/MS and antimicrobial disk diffusion susceptibility testing for positive blood cultures after a short incubation on the WASPLab

Authors: Abdessalam Cherkaoui, Gesuele Renzi, Nouria Azam, Didier Schorderet, Nicolas Vuilleumier, Jacques Schrenzel

Published in: European Journal of Clinical Microbiology & Infectious Diseases | Issue 6/2020

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Abstract

The objectives of this study were to define the shortest incubation times on the WASPLab for reliable MALDI-TOF/MS-based species identification and for the preparation of a 0.5 McFarland suspension for antimicrobial disk diffusion susceptibility testing using short subcultures growing on solid culture media inoculated by positive blood cultures spiked with a wide range of pathogens associated with bloodstream infections. The 520 clinical strains (20 × 26 different species) included in this study were obtained from a collection of non-consecutive and non-duplicate pathogens identified at Geneva University Hospitals. After 4 h of incubation on the WASPLab, microorganisms’ growth allowed accurate identification of 73% (380/520) (95% CI, 69.1–76.7%) of the strains included in this study. The identification rate increased to 85% (440/520) (95% CI, 81.3–87.5%) after 6-h incubation. When excluding Corynebacterium and Candida spp., the microbial growth was sufficient to permit accurate identification of all tested species (100%, 460/460) (95% CI, 99.2–100%) after 8-h incubation. With the exception of Burkholderia cepacia and Haemophilus influenzae, AST by disk diffusion could be performed for Enterobacterales and non-fermenting Gram-negative bacilli after only 4 h of growth in the WASPLab. The preparation of a 0.5 McFarland suspension for Gram-positive bacteria required incubation times ranging between 3 and 8 h according to the bacterial species. Only Corynebacterium spp. required incubation times as long as 16 h. The WASPLab enables rapid pathogen identification as well as swift comprehensive AST from positive blood cultures that can be implemented without additional costs nor hands-on time by defining optimal time points for image acquisition.
Literature
1.
go back to reference Gaieski DF, Mikkelsen ME, Band RA, Pines JM, Massone R, Furia FF, Shofer FS, Goyal M (2010) Impact of time to antibiotics on survival in patients with severe sepsis or septic shock in whom early goal-directed therapy was initiated in the emergency department. Crit Care Med 38(4):1045–1053CrossRef Gaieski DF, Mikkelsen ME, Band RA, Pines JM, Massone R, Furia FF, Shofer FS, Goyal M (2010) Impact of time to antibiotics on survival in patients with severe sepsis or septic shock in whom early goal-directed therapy was initiated in the emergency department. Crit Care Med 38(4):1045–1053CrossRef
2.
go back to reference Larche J, Azoulay E, Fieux F, Mesnard L, Moreau D, Thiery G, Darmon M, Le Gall JR, Schlemmer B (2003) Improved survival of critically ill cancer patients with septic shock. Intensive Care Med 29(10):1688–1695CrossRef Larche J, Azoulay E, Fieux F, Mesnard L, Moreau D, Thiery G, Darmon M, Le Gall JR, Schlemmer B (2003) Improved survival of critically ill cancer patients with septic shock. Intensive Care Med 29(10):1688–1695CrossRef
3.
go back to reference Garnacho-Montero J, Aldabo-Pallas T, Garnacho-Montero C, Cayuela A, Jimenez R, Barroso S, Ortiz-Leyba C (2006) Timing of adequate antibiotic therapy is a greater determinant of outcome than are TNF and IL-10 polymorphisms in patients with sepsis. Crit Care 10(4):R111CrossRef Garnacho-Montero J, Aldabo-Pallas T, Garnacho-Montero C, Cayuela A, Jimenez R, Barroso S, Ortiz-Leyba C (2006) Timing of adequate antibiotic therapy is a greater determinant of outcome than are TNF and IL-10 polymorphisms in patients with sepsis. Crit Care 10(4):R111CrossRef
4.
go back to reference Kumar A, Roberts D, Wood KE, Light B, Parrillo JE, Sharma S, Suppes R, Feinstein D, Zanotti S, Taiberg L, Gurka D, Kumar A, Cheang M (2006) Duration of hypotension before initiation of effective antimicrobial therapy is the critical determinant of survival in human septic shock. Crit Care Med 34(6):1589–1596CrossRef Kumar A, Roberts D, Wood KE, Light B, Parrillo JE, Sharma S, Suppes R, Feinstein D, Zanotti S, Taiberg L, Gurka D, Kumar A, Cheang M (2006) Duration of hypotension before initiation of effective antimicrobial therapy is the critical determinant of survival in human septic shock. Crit Care Med 34(6):1589–1596CrossRef
5.
go back to reference Leal HF, Azevedo J, Silva GEO, Amorim AML, de Roma LRC, Arraes ACP, Gouveia EL, Reis MG, Mendes AV, de Oliveira SM, Barberino MG, Martins IS, Reis JN (2019) Bloodstream infections caused by multidrug-resistant gram-negative bacteria: epidemiological, clinical and microbiological features. BMC Infect Dis 19(1):609CrossRef Leal HF, Azevedo J, Silva GEO, Amorim AML, de Roma LRC, Arraes ACP, Gouveia EL, Reis MG, Mendes AV, de Oliveira SM, Barberino MG, Martins IS, Reis JN (2019) Bloodstream infections caused by multidrug-resistant gram-negative bacteria: epidemiological, clinical and microbiological features. BMC Infect Dis 19(1):609CrossRef
6.
go back to reference Matos ECO, Andriolo RB, Rodrigues YC, Lima PDL, Carneiro I, Lima KVB (2018) Mortality in patients with multidrug-resistant Pseudomonas aeruginosa infections: a meta-analysis. Rev Soc Bras Med Trop 51(4):415–420CrossRef Matos ECO, Andriolo RB, Rodrigues YC, Lima PDL, Carneiro I, Lima KVB (2018) Mortality in patients with multidrug-resistant Pseudomonas aeruginosa infections: a meta-analysis. Rev Soc Bras Med Trop 51(4):415–420CrossRef
7.
go back to reference Liesenfeld O, Lehman L, Hunfeld KP, Kost G (2014) Molecular diagnosis of sepsis: new aspects and recent developments. Eur J Microbiol Immunol (Bp) 4(1):1–25CrossRef Liesenfeld O, Lehman L, Hunfeld KP, Kost G (2014) Molecular diagnosis of sepsis: new aspects and recent developments. Eur J Microbiol Immunol (Bp) 4(1):1–25CrossRef
8.
go back to reference Candel FJ, Borges Sa M, Belda S, Bou G, Del Pozo JL, Estrada O, Ferrer R, Gonzalez Del Castillo J, Julian-Jimenez A, Martin-Loeches I, Maseda E, Matesanz M, Ramirez P, Ramos JT, Rello J, Suberviola B, Suarez de la Rica A, Vidal P (2018) Current aspects in sepsis approach. Turning things around. Rev Esp Quimioter 31(4):298–315PubMedPubMedCentral Candel FJ, Borges Sa M, Belda S, Bou G, Del Pozo JL, Estrada O, Ferrer R, Gonzalez Del Castillo J, Julian-Jimenez A, Martin-Loeches I, Maseda E, Matesanz M, Ramirez P, Ramos JT, Rello J, Suberviola B, Suarez de la Rica A, Vidal P (2018) Current aspects in sepsis approach. Turning things around. Rev Esp Quimioter 31(4):298–315PubMedPubMedCentral
9.
go back to reference Cherkaoui A, Hibbs J, Emonet S, Tangomo M, Girard M, Francois P, Schrenzel J (2010) Comparison of two matrix-assisted laser desorption ionization-time of flight mass spectrometry methods with conventional phenotypic identification for routine identification of bacteria to the species level. J Clin Microbiol 48(4):1169–1175CrossRef Cherkaoui A, Hibbs J, Emonet S, Tangomo M, Girard M, Francois P, Schrenzel J (2010) Comparison of two matrix-assisted laser desorption ionization-time of flight mass spectrometry methods with conventional phenotypic identification for routine identification of bacteria to the species level. J Clin Microbiol 48(4):1169–1175CrossRef
10.
go back to reference Ferreira L, Sanchez-Juanes F, Porras-Guerra I, Garcia-Garcia MI, Garcia-Sanchez JE, Gonzalez-Buitrago JM, Munoz-Bellido JL (2011) Microorganisms direct identification from blood culture by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Clin Microbiol Infect 17(4):546–551CrossRef Ferreira L, Sanchez-Juanes F, Porras-Guerra I, Garcia-Garcia MI, Garcia-Sanchez JE, Gonzalez-Buitrago JM, Munoz-Bellido JL (2011) Microorganisms direct identification from blood culture by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Clin Microbiol Infect 17(4):546–551CrossRef
11.
go back to reference Christner M, Rohde H, Wolters M, Sobottka I, Wegscheider K, Aepfelbacher M (2010) 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 48(5):1584–1591CrossRef Christner M, Rohde H, Wolters M, Sobottka I, Wegscheider K, Aepfelbacher M (2010) 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 48(5):1584–1591CrossRef
12.
go back to reference Haigh JD, Green IM, Ball D, Eydmann M, Millar M, Wilks M (2013) Rapid identification of bacteria from bioMerieux BacT/ALERT blood culture bottles by MALDI-TOF MS. Br J Biomed Sci 70(4):149–155CrossRef Haigh JD, Green IM, Ball D, Eydmann M, Millar M, Wilks M (2013) Rapid identification of bacteria from bioMerieux BacT/ALERT blood culture bottles by MALDI-TOF MS. Br J Biomed Sci 70(4):149–155CrossRef
13.
go back to reference Campigotto A, Goneau L, Matukas LM (2018) Direct identification and antimicrobial susceptibility testing of microorganisms from positive blood cultures following isolation by lysis-centrifugation. Diagn Microbiol Infect Dis 92(3):189–193CrossRef Campigotto A, Goneau L, Matukas LM (2018) Direct identification and antimicrobial susceptibility testing of microorganisms from positive blood cultures following isolation by lysis-centrifugation. Diagn Microbiol Infect Dis 92(3):189–193CrossRef
14.
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(Pt 11):1511–1516CrossRef 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(Pt 11):1511–1516CrossRef
15.
go back to reference Klein S, Zimmermann S, Kohler C, Mischnik A, Alle W, Bode KA (2012) Integration of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry in blood culture diagnostics: a fast and effective approach. J Med Microbiol 61(Pt 3):323–331CrossRef Klein S, Zimmermann S, Kohler C, Mischnik A, Alle W, Bode KA (2012) Integration of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry in blood culture diagnostics: a fast and effective approach. J Med Microbiol 61(Pt 3):323–331CrossRef
16.
go back to reference Kohlmann R, Hoffmann A, Geis G, Gatermann S (2015) 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 305(4–5):469–479CrossRef Kohlmann R, Hoffmann A, Geis G, Gatermann S (2015) 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 305(4–5):469–479CrossRef
17.
go back to reference Thomin J, Aubin GG, Foubert F, Corvec S (2015) Assessment of four protocols for rapid bacterial identification from positive blood culture pellets by matrix-assisted laser desorption ionization-time of flight mass spectrometry (Vitek(R) MS). J Microbiol Methods 115:54–56CrossRef Thomin J, Aubin GG, Foubert F, Corvec S (2015) Assessment of four protocols for rapid bacterial identification from positive blood culture pellets by matrix-assisted laser desorption ionization-time of flight mass spectrometry (Vitek(R) MS). J Microbiol Methods 115:54–56CrossRef
18.
go back to reference March GA, Garcia-Loygorri MC, Simarro M, Gutierrez MP, Orduna A, Bratos MA (2015) A new approach to determine the susceptibility of bacteria to antibiotics directly from positive blood culture bottles in two hours. J Microbiol Methods 109:49–55CrossRef March GA, Garcia-Loygorri MC, Simarro M, Gutierrez MP, Orduna A, Bratos MA (2015) A new approach to determine the susceptibility of bacteria to antibiotics directly from positive blood culture bottles in two hours. J Microbiol Methods 109:49–55CrossRef
19.
go back to reference Jakovljev A, Bergh K (2015) Development of a rapid and simplified protocol for direct bacterial identification from positive blood cultures by using matrix assisted laser desorption ionization time-of- flight mass spectrometry. BMC Microbiol 15:258CrossRef Jakovljev A, Bergh K (2015) Development of a rapid and simplified protocol for direct bacterial identification from positive blood cultures by using matrix assisted laser desorption ionization time-of- flight mass spectrometry. BMC Microbiol 15:258CrossRef
20.
go back to reference Fitzgerald C, Stapleton P, Phelan E, Mulhare P, Carey B, Hickey M, Lynch B (2016 Apr) Doyle M (2016) rapid identification and antimicrobial susceptibility testing of positive blood cultures using MALDI-TOF MS and a modification of the standardised disc diffusion test: a pilot study. J Clin Pathol 27 Fitzgerald C, Stapleton P, Phelan E, Mulhare P, Carey B, Hickey M, Lynch B (2016 Apr) Doyle M (2016) rapid identification and antimicrobial susceptibility testing of positive blood cultures using MALDI-TOF MS and a modification of the standardised disc diffusion test: a pilot study. J Clin Pathol 27
21.
go back to reference Verroken A, Defourny L, le Polain de Waroux O, Belkhir L, Laterre PF, Delmee M, Glupczynski Y (2016) Clinical impact of MALDI-TOF MS identification and rapid susceptibility testing on adequate antimicrobial treatment in Sepsis with positive blood cultures. PLoS One 11(5):e0156299CrossRef Verroken A, Defourny L, le Polain de Waroux O, Belkhir L, Laterre PF, Delmee M, Glupczynski Y (2016) Clinical impact of MALDI-TOF MS identification and rapid susceptibility testing on adequate antimicrobial treatment in Sepsis with positive blood cultures. PLoS One 11(5):e0156299CrossRef
22.
go back to reference Kim JM, Kim I, Chung SH, Chung Y, Han M, Kim JS (2019) Rapid discrimination of methicillin-resistant Staphylococcus aureus by MALDI-TOF MS. Pathogens 8(4) Kim JM, Kim I, Chung SH, Chung Y, Han M, Kim JS (2019) Rapid discrimination of methicillin-resistant Staphylococcus aureus by MALDI-TOF MS. Pathogens 8(4)
23.
go back to reference Axelsson C, Rehnstam-Holm AS, Nilson B (2019) Rapid detection of antibiotic resistance in positive blood cultures by MALDI-TOF MS and an automated and optimized MBT-ASTRA protocol for Escherichia coli and Klebsiella pneumoniae. Infect Dis (Lond):1–9 Axelsson C, Rehnstam-Holm AS, Nilson B (2019) Rapid detection of antibiotic resistance in positive blood cultures by MALDI-TOF MS and an automated and optimized MBT-ASTRA protocol for Escherichia coli and Klebsiella pneumoniae. Infect Dis (Lond):1–9
24.
go back to reference Cherkaoui A, Cherpillod P, Renzi G, Schrenzel J, Kaiser L, Schibler M (2019) A molecular based diagnosis of positive blood culture in the context of viral haemorrhagic fever: proof of concept. Clin Microbiol Infect 25(10):1289 e1281–1289 e1284CrossRef Cherkaoui A, Cherpillod P, Renzi G, Schrenzel J, Kaiser L, Schibler M (2019) A molecular based diagnosis of positive blood culture in the context of viral haemorrhagic fever: proof of concept. Clin Microbiol Infect 25(10):1289 e1281–1289 e1284CrossRef
25.
go back to reference Huang TD, Melnik E, Bogaerts P, Evrard S, Glupczynski Y (2019) Evaluation of the ePlex blood culture identification panels for detection of pathogens in bloodstream infections. J Clin Microbiol 57(2) Huang TD, Melnik E, Bogaerts P, Evrard S, Glupczynski Y (2019) Evaluation of the ePlex blood culture identification panels for detection of pathogens in bloodstream infections. J Clin Microbiol 57(2)
26.
go back to reference Kolb M, Lazarevic V, Emonet S, Calmy A, Girard M, Gaia N, Charretier Y, Cherkaoui A, Keller P, Huber C, Schrenzel J (2019) Next-generation sequencing for the diagnosis of challenging culture-negative endocarditis. Front Med (Lausanne) 6:203CrossRef Kolb M, Lazarevic V, Emonet S, Calmy A, Girard M, Gaia N, Charretier Y, Cherkaoui A, Keller P, Huber C, Schrenzel J (2019) Next-generation sequencing for the diagnosis of challenging culture-negative endocarditis. Front Med (Lausanne) 6:203CrossRef
27.
go back to reference Lazarevic V, Gaia N, Girard M, Leo S, Cherkaoui A, Renzi G, Emonet S, Jamme S, Ruppe E, Vijgen S, Rubbia-Brandt L, Toso C, Schrenzel J (2018) When bacterial culture fails, Metagenomics can help: a case of chronic hepatic Brucelloma assessed by next-generation sequencing. Front Microbiol 9:1566CrossRef Lazarevic V, Gaia N, Girard M, Leo S, Cherkaoui A, Renzi G, Emonet S, Jamme S, Ruppe E, Vijgen S, Rubbia-Brandt L, Toso C, Schrenzel J (2018) When bacterial culture fails, Metagenomics can help: a case of chronic hepatic Brucelloma assessed by next-generation sequencing. Front Microbiol 9:1566CrossRef
Metadata
Title
Rapid identification by MALDI-TOF/MS and antimicrobial disk diffusion susceptibility testing for positive blood cultures after a short incubation on the WASPLab
Authors
Abdessalam Cherkaoui
Gesuele Renzi
Nouria Azam
Didier Schorderet
Nicolas Vuilleumier
Jacques Schrenzel
Publication date
01-06-2020
Publisher
Springer Berlin Heidelberg
Published in
European Journal of Clinical Microbiology & Infectious Diseases / Issue 6/2020
Print ISSN: 0934-9723
Electronic ISSN: 1435-4373
DOI
https://doi.org/10.1007/s10096-020-03817-8

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