Proteome-based bacterial identification using matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS): A revolutionary shift in clinical diagnostic microbiology

Nomura, Fumio

doi:10.1016/j.bbapap.2014.10.022

Cited by 113 publications

(86 citation statements)

References 84 publications

(93 reference statements)

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“…Diagnoses of clinical brucellosis are made initially using serological tests and are confirmed by isolation of the agent (6,7). The recent introduction of matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry (MS) has revolutionized the identification of bacteria and yeasts (8,9). However, the procedures recommended by the manufacturers for non-biosafety level 3 (BSL3) organisms are not adequate for the manipulation of Brucella strains, which are classified as BSL3 (potential bioterrorism pathogens) and represent potential health hazards for laboratory workers (1,10).…”

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confidence: 99%

A Simple and Safe Protocol for Preparing Brucella Samples for Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry Analysis

et al. 2016

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dWe describe a simple protocol to inactivate the biosafety level 3 (BSL3) pathogens Brucella prior to their analysis by matrix-assisted laser desorption ionization-time of flight mass spectrometry. This method is also effective for several other bacterial pathogens and allows storage, and eventually shipping, of inactivated samples; therefore, it might be routinely applied to unidentified bacteria, for the safety of laboratory workers. Brucellosis is a zoonosis caused by bacteria of the genus Brucella, which are transmitted to humans through direct contact, ingestion of contaminated animal products, or aerosolization (1, 2). Brucellosis is endemic in several regions of the world, with more than 500,000 new cases each year (2-5). To date, 10 recognized species of Brucella have been described, with the most pathogenic for humans being Brucella melitensis (2). Diagnoses of clinical brucellosis are made initially using serological tests and are confirmed by isolation of the agent (6, 7). The recent introduction of matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry (MS) has revolutionized the identification of bacteria and yeasts (8, 9). However, the procedures recommended by the manufacturers for non-biosafety level 3 (BSL3) organisms are not adequate for the manipulation of Brucella strains, which are classified as BSL3 (potential bioterrorism pathogens) and represent potential health hazards for laboratory workers (1, 10). Any inactivation procedure must allow safe sample handling outside the BSL3 environment and must avoid destruction of the biomarkers used for identification. Therefore, we developed a simple, safe, and efficient sample preparation method for Brucella isolates that is compatible with their analysis by the current MALDI-TOF MS platforms.Solvent inactivation of Brucella. We first applied the direct transfer procedure recommended by the manufacturer to a panel of Brucella strains, working in class II microbiological safety cabinets in a BSL3 laboratory (Table 1). The target plate (Vitek MS DS slide; bioMérieux) was inoculated by picking a portion of a colony from a plate with a 1-l disposable loop (Sarsted), and the deposit was overlaid with 1 l of ␣-cyano-4-hydroxycinnamic acid (CHCA) matrix solution (saturated solution of CHCA in a solvent mixture composed of 33% acetonitrile, 33% absolute ethanol, 3% trifluoroacetic acid, and 31% water) and then air dried for 2 min at room temperature. The dried material was recovered with a sterile swab and spread on culture plates, which were then incubated for up to 3 weeks under the optimal growth conditions for the corresponding bacteria indicated in Table 1. Viable bacteria were recovered for most strains, showing that this protocol does not kill all Brucella strains, possibly because the bacterial deposit was not completely covered with the matrix solution. This risk is not acceptable for BSL3 pathogens. To overcome this problem, different liquid-phase inactivation protocols were tested in 1.5-ml Eppendorf tubes (see F...

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A Simple and Safe Protocol for Preparing Brucella Samples for Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry Analysis

et al. 2016

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“…24 The spectra that are obtained represent a unique fingerprint for each microorganism and are compared with a library of spectra of known species to make the identification. 25,26 Thus this technology is ideal for an accurate fungal identification at the species level, including differentiation of closely related species and subspecies. Mass spectrometry was first used in microbial testing in 1975, when bacteria with unique mass spectra signature were characterized for the first time.…”

Section: Matrix-assisted Laser Desorption/ionization Time-of-flight Mmentioning

confidence: 99%

Update from the Laboratory

Bataineh

Sutton

Fothergill

et al. 2016

Infectious Disease Clinics of North America

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“…In particular, matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI)-TOF has proven to be a fast and reliable method for identification of bacteria in clinical samples [83]. That said, examples of proteomic-based pathogen identification in neurological infections are just emerging.…”

Section: Proteomics Studies On Bacterial Infections Of the Nervous Symentioning

confidence: 99%

Application of “Omics” Technologies for Diagnosis and Pathogenesis of Neurological Infections

Noorbakhsh

Aminian

Power

2015

Curr Neurol Neurosci Rep

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Infections of the human nervous system have substantial morbidity and mortality but also represent among the most challenging of all neurological diseases because of the difficulty in establishing a diagnosis and implementing effective therapies. Neurological infections lead to altered expression levels of a wide range of host- and pathogen-derived biomolecules both within and outside of the nervous system. Quantitative analyses of these biomolecular perturbations have been traditionally performed using "classical" molecular or analytical methods, which evaluate one or few genes or their products at a time. Recent technical developments together with the increasing availability of high-throughput/content methodologies have enabled a more comprehensive overview of these molecular alterations and thus provide new approaches to the diagnosis and/or treatment of this group of disorders. Herein, we will review recent evidence pointing to the capacity of the so-called omics techniques in studying the nervous system infections with an emphasis on genomics, transcriptomics, and proteomics technologies.

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Proteome-based bacterial identification using matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS): A revolutionary shift in clinical diagnostic microbiology

Cited by 113 publications

References 84 publications

A Simple and Safe Protocol for Preparing Brucella Samples for Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry Analysis

A Simple and Safe Protocol for Preparing Brucella Samples for Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry Analysis

Update from the Laboratory

Application of “Omics” Technologies for Diagnosis and Pathogenesis of Neurological Infections

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