Matrix-Assisted Laser Desorption Ionization–Time of Flight (MALDI-TOF) Mass Spectrometry for Detection of Antibiotic Resistance Mechanisms: from Research to Routine Diagnosis

Hrabák, Jaroslav; Chudáčková, Eva; Walková, Radka

doi:10.1128/cmr.00058-12

Cited by 249 publications

(168 citation statements)

References 78 publications

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“…Although in its early stages, several publications are summarized and describe the successful use of MALDI-TOF MS for discriminating antibiotic-resistant bacterial strains. While a comprehensive description of the use of MALDI-TOF MS for the identification of microbes in the clinical laboratory is expertly reviewed elsewhere (324) and is outside the scope of this review, we will instead highlight some of the uses of MALDI-TOF MS for the detection of different groups of clinically important antibiotic-resistant bacteria.…”

Section: Detection Of Resistance To Beta-lactam Antibiotics In Enterimentioning

confidence: 99%

Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry: a Fundamental Shift in the Routine Practice of Clinical Microbiology

et al. 2013

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SUMMARY Within the past decade, clinical microbiology laboratories experienced revolutionary changes in the way in which microorganisms are identified, moving away from slow, traditional microbial identification algorithms toward rapid molecular methods and mass spectrometry (MS). Historically, MS was clinically utilized as a high-complexity method adapted for protein-centered analysis of samples in chemistry and hematology laboratories. Today, matrix-assisted laser desorption ionization–time of flight (MALDI-TOF) MS is adapted for use in microbiology laboratories, where it serves as a paradigm-shifting, rapid, and robust method for accurate microbial identification. Multiple instrument platforms, marketed by well-established manufacturers, are beginning to displace automated phenotypic identification instruments and in some cases genetic sequence-based identification practices. This review summarizes the current position of MALDI-TOF MS in clinical research and in diagnostic clinical microbiology laboratories and serves as a primer to examine the “nuts and bolts” of MALDI-TOF MS, highlighting research associated with sample preparation, spectral analysis, and accuracy. Currently available MALDI-TOF MS hardware and software platforms that support the use of MALDI-TOF with direct and precultured specimens and integration of the technology into the laboratory workflow are also discussed. Finally, this review closes with a prospective view of the future of MALDI-TOF MS in the clinical microbiology laboratory to accelerate diagnosis and microbial identification to improve patient care.

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Section: Detection Of Resistance To Beta-lactam Antibiotics In Enterimentioning

confidence: 99%

Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry: a Fundamental Shift in the Routine Practice of Clinical Microbiology

et al. 2013

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“…Hence, microbiologists should be regularly updated with regional surveillance data to ensure that state-of-the art screening and confirmatory testing are in place (187,188). Molecular and protein-based identification tools should also be considered to improve diagnosis and to reduce turnaround time (189).…”

Section: The Need For Regional Surveillance Studiesmentioning

confidence: 99%

β-Lactamase Production in Key Gram-Negative Pathogen Isolates from the Arabian Peninsula

et al. 2013

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SUMMARY Infections due to Gram-negative bacilli (GNB) are a leading cause of morbidity and mortality worldwide. The extent of antibiotic resistance in GNB in countries of the Gulf Cooperation Council (GCC), namely, Saudi Arabia, United Arab Emirates, Kuwait, Qatar, Oman, and Bahrain, has not been previously reviewed. These countries share a high prevalence of extended-spectrum-β-lactamase (ESBL)- and carbapenemase-producing GNB, most of which are associated with nosocomial infections. Well-known and widespread β-lactamases genes (such as those for CTX-M-15, OXA-48, and NDM-1) have found their way into isolates from the GCC states. However, less common and unique enzymes have also been identified. These include PER-7, GES-11, and PME-1. Several potential risk factors unique to the GCC states may have contributed to the emergence and spread of β-lactamases, including the unnecessary use of antibiotics and the large population of migrant workers, particularly from the Indian subcontinent. It is clear that active surveillance of antimicrobial resistance in the GCC states is urgently needed to address regional interventions that can contain the antimicrobial resistance issue.

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“…Detection of ␤-lactamases by MALDI-TOF MS is a proteomic approach allowing the study of the behavior of the tested strains and should complement techniques already used for characterization of ␤-lactamases such as PCR and isoelectric focusing (IEF). The fact that MALDI-TOF MS can directly detect class A (9) and class C ␤-lactamases, as well as other mechanisms such as methylation of rRNA and cell wall components (3,27), indicates the feasibility of establishing a MALDI-TOF supplementary database of resistance mechanisms that would promote research in this field. Notwithstanding the aforementioned problems, we strongly believe that proper modifications and validation of the described MALDI-TOF assay will easily lead to acceptance of its future application in diagnostic laboratories and reference centers.…”

Section: -245mentioning

confidence: 99%

“…Recently, further applications of MALDI-TOF MS focusing on antimicrobial resistance mechanisms, including detection of carbapenemase activity in Enterobacteriaceae, Pseudomonas spp., and Acinetobacter spp., have been described (3)(4)(5)(6)(7).…”

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confidence: 99%

Identification of CMY-2-Type Cephalosporinases in Clinical Isolates of Enterobacteriaceae by MALDI-TOF MS

Papagiannitsis

Kotsakis

Tůma

et al. 2014

Antimicrob Agents Chemother

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This study exploited the possibility to detect Citrobacter freundii-derived CMY-2-like cephalosporinases in Enterobacteriaceae clinical isolates using matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). Periplasmic proteins were prepared using a modified sucrose method and analyzed by MALDI-TOF MS. A ca. 39,850-m/z peak, confirmed to represent a C. freundii-like ␤-lactamase by in-gel tryptic digestion followed by MALDI-TOF/TOF MS, was observed only in CMY-producing isolates. We have also shown the potential of the assay to detect ACC-and DHA-like AmpC-type ␤-lactamases. Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) is increasingly used as a procedure for identification of pathogenic bacteria and fungi due to its time-and cost-effectiveness (1, 2). Recently, further applications of MALDI-TOF MS focusing on antimicrobial resistance mechanisms, including detection of carbapenemase activity in Enterobacteriaceae, Pseudomonas spp., and Acinetobacter spp., have been described (3-7).In 2007, Camara and Hays described for the first time the use of MALDI-TOF MS for differentiating wild-type Escherichia coli from ampicillin-resistant (Amp r ) plasmid-transformed E. coli strains by the direct visualization of a ␤-lactamase (8). In a recent MALDI-TOF MS study, Schaumann et al. were not able to distinguish Enterobacteriaceae and P. aeruginosa isolates producing extended-spectrum ␤-lactamases (ESBLs) or metallo-␤-lactamases (MBLs) from nonproducers (9). Consequently, so far the attempts to visualize native ␤-lactamases by MALDI-TOF MS in wild-type bacteria have been mostly unsuccessful.We describe here a new assay for the identification of CMY-2-like ␤-lactamases in clinical enterobacterial isolates by MALDI-TOF MS. These enzymes are the most prevalent acquired AmpCtype cephalosporinases in Enterobacteriaceae (10). The method is based on the extraction of periplasmic proteins and the detection of CMY-2-like ␤-lactamases by MALDI-TOF MS according to their molecular weight.Thirty-eight characterized Enterobacteriaceae strains from collections of the Faculty of Medicine and University Hospital in Plzen, Czech Republic, the Hellenic Pasteur Institute in Athens, Greece, and the National Medicines Institute in Warsaw, Poland, were used (Table 1) (11,12). The group included 29 CMY-2-positive clinical isolates, two E. coli transconjugants/transformants with CMY-2-like enzymes (E. coli A15 or DH5␣), and seven non-CMY-producing isolates (13-21). E. coli ATCC 25922 and Klebsiella pneumoniae ATCC 13883 were used as negative controls. Purified CMY-2 enzyme (13) was used as a positive control for MALDI-TOF MS measurements.Isolates were inoculated into 50 ml of Mueller-Hinton broth (Oxoid Ltd.) and incubated at 35°C for 16 h. Cultures were centrifuged at 5,000 ϫ g for 20 min, and the cell pellet was used for the extraction of the periplasmic proteins, performed essentially as described by Naglak and Wang (22). Briefly, the pellet was resuspended in 360 l ...

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Matrix-Assisted Laser Desorption Ionization–Time of Flight (MALDI-TOF) Mass Spectrometry for Detection of Antibiotic Resistance Mechanisms: from Research to Routine Diagnosis

Cited by 249 publications

References 78 publications

Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry: a Fundamental Shift in the Routine Practice of Clinical Microbiology

Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry: a Fundamental Shift in the Routine Practice of Clinical Microbiology

β-Lactamase Production in Key Gram-Negative Pathogen Isolates from the Arabian Peninsula

Identification of CMY-2-Type Cephalosporinases in Clinical Isolates of Enterobacteriaceae by MALDI-TOF MS

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