Fast and reliable bacterial identification direct from positive blood culture using a new TFA sample preparation protocol and the Vitek® MS system

Monteiro, Jussimara; Inoue, Fernanda Matsiko; Lobo, Ana Paula Timm; Sugawara, Eduardo Kinio; Boaretti, Fabiana M.; Tufik, Sérgio

doi:10.1016/j.mimet.2014.12.009

Cited by 26 publications

(22 citation statements)

References 10 publications

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“…The composition of Gram-negative (GN) cell walls is such that extraction of proteins through common MALDI-TOF MS protocols is reliable and highly accurate. Purification methods from positive blood culture have results similar to those of direct colony testing, with concordance ranging from 84 to 99% to a species-level identification, with most studies observing Ͼ90% concordance (19,20). In one study, the authors found reliable identification to the genus (but not species) level for Enterobacter cloacae and to the species level for Proteus vulgaris (n ϭ 5 for each organism) (16).…”

Section: Assay Performance and Optimizationmentioning

confidence: 95%

“…Low-speed centrifugation is used to remove the erythrocytes from the bacterial cells, often followed by an additional lysis procedure to further remove any remnant blood cells. Common reagents for lysis include 5% saponin, ammonium chloride, trifluoroacetic acid (TFA), 10ϫ Triton, and sterile water (16)(17)(18)(19)(20)(21)(22). Lysed samples are then centrifuged to isolate a bacterial pellet and washed with either water, ethanol, or a saline solution.…”

Section: Methodsmentioning

confidence: 99%

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Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry for Use with Positive Blood Cultures: Methodology, Performance, and Optimization

2017

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Early initiation of effective antibiotics for septic patients is essential for patient survival. Matrix-assisted desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) has revolutionized clinical microbiology for isolate identification and has the possibility to impact how blood culture testing is performed. This review discusses the various uses of MALDI-TOF MS for the identification and susceptibility testing of positive blood cultures, the performance of these methods, and the outcomes involved with its implementation.

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Section: Assay Performance and Optimizationmentioning

confidence: 95%

Section: Methodsmentioning

confidence: 99%

Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry for Use with Positive Blood Cultures: Methodology, Performance, and Optimization

2017

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“…An American study compared the BD Phoenix automated microbiology system to MALDI-TOF MS for Gram-negative bacilli identification (MONTEIRO et al, 2015). MicroFlex LT system was used to MALDI-TOF MS and the spectra were analyzed using Bruker Biotyper 2.0 software.…”

Section: Maldi-tof Ms Data Analysismentioning

confidence: 99%

“…Numerous publications reported MALDI-TOF MS applications on identification of diverse bacterial groups, genus or species such as Enterobacteriaceae (BIBERG et al, 2015), non-fermenting bacteria (MELLMANN et al, 2008), Staphylococcus spp. (DUBOIS et al, 2010), Neisseria (ILINA et al, 2009, Clostridia (GROSSE-HERRENTHEY et al, 2008), Mycobacteria (PIGNONE et al, 2006), Helicobacter pylori (PEREIRA et al, 2006), Campylobacter (MARTINY et al, 2011), Mycoplasma hominis (PAILHORIÈS et al, 2014) and some Haemophilus species (FRICKMANN et al, 2013).…”

Section: Maldi-tof Ms Performance In Bacterial Identificationmentioning

confidence: 99%

Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI-TOF MS) Applications in Bacteriology: brazilian contributions

Pellegrino

Chagas

Alves

et al. 2018

HU Rev

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Among its innumerous applications in Bacteriology, the Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI-TOF MS) technique is evolving as a powerful tool for bacterial identification and antimicrobial resistance investigation. Publications have evaluated the MALDI-TOF MS performance in the identification of a series of bacterial pathogens, including the most common severe infectious agents, emergent pathogens involved with outbreaks of healthcare-associated infections, rare pathogens, and those whose isolation in culture media is difficult. As compared to conventional methods of bacterial identification, MALDI-TOF MS has proven to be a fast, accurate and cost-effective technique. Currently, MALDI-TOF MS has been used in antimicrobial resistance studies, since it has shown to be an efficient tool in detecting specific resistance mechanisms in bacteria, such as beta-lactamases production, for example. Here, we describe the advances in this growing field of mass spectrometry applied to Bacteriology, including Brazilian contributions.

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“…Therefore, lysis solutions [22] , the lysis-filtration method [12] or commercial kits [19] were utilized for sample preparation. However, some of these procedures are laborious and/or expensive [23] .…”

Section: Introductionmentioning

confidence: 99%

Identification and antimicrobial susceptibility testing of microorganisms from positive blood cultures by a combined lysis-centrifugation method with MALDI-TOF MS and VITEK2 System

Rodio

Febbraro

Puggioni

et al. 2017

APALM

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Background: Rapid identification and the application of antimicrobial susceptibility testing (AST) to microorganisms causing bloodstream infections is pivotal to guide antimicrobial therapy. This study aims to: 1) utilize the Lysis-Centrifugation Method (LCM) not only for identification of microorganisms from positive blood culture bottles, but also for direct AST full panel by Vitek ® 2 system (bioMérieux, Inc. France) and by disc diffusion plate (Kirby Bauer Method) and 2) analyze the accuracy of these combined methods.Methods: 124 mono-microbial positive blood culture bottles were included in this study. An aliquot was subjected to LCM and used for the identification by the MALDI-TOF System. Moreover the microbial pellet was used for direct AST testing full panel by VITEK Results: 123 isolates were correctly identified to the species level and 1 isolate was identified to the genus level. Comparing the two utilized AST methods, it was observed that Gram-positive isolates showed an agreement rate of 96.6% (58/60). Enterococcus faecalis was the only microorganism with a major error rate of 0.6% (2/324) related to erythromycin. Among the Gram-negative, the overall agreement rate was 93.3 (56/60). Klebsiella pneumoniae, Escherichia coli and Enterobacter spp. were the major cause of minor error rates (0.6%, 4/709) and major error rates (1.1%, 8/709). Among the yeasts, results showed an agreement rate of 100% (4/4). Conclusions:Our simple and cost-effective sample preparation method is very useful for rapid identification as well as AST of microorganisms directly from positive blood culture bottles in a clinical setting.

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Fast and reliable bacterial identification direct from positive blood culture using a new TFA sample preparation protocol and the Vitek® MS system

Cited by 26 publications

References 10 publications

Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry for Use with Positive Blood Cultures: Methodology, Performance, and Optimization

Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry for Use with Positive Blood Cultures: Methodology, Performance, and Optimization

Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI-TOF MS) Applications in Bacteriology: brazilian contributions

Identification and antimicrobial susceptibility testing of microorganisms from positive blood cultures by a combined lysis-centrifugation method with MALDI-TOF MS and VITEK2 System

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