Biotyping of Multidrug-Resistant Klebsiella pneumoniae Clinical Isolates from France and Algeria Using MALDI-TOF MS

Berrazeg, Meryem; Diène, Seydina M.; Drissi, Mourad; Kempf, Marie; Richet, Hervé; Landraud, Luce; Rolain, Jean‐Marc

doi:10.1371/journal.pone.0061428

Cited by 76 publications

(57 citation statements)

References 37 publications

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“…Furthermore, it is a powerful method and is sufficiently reproducible and sensitive enough to rapidly survey the evolution of existing or emerging phenotypes with reduced financial and human costs (4 In the current study, the bioMérieux VITEK MS system was able to identify the Klebsiella species with an accuracy of 100%, and the reliability of some results was confirmed by sequencing analysis. Interest in the VITEK 2 automated system has increased due to its minimization of errors, such as indicating false susceptibility for resistant microorganisms, the computer included in its system, its reduction of workload, and its objectivity in evaluation of the results.…”

Section: Discussionmentioning

confidence: 57%

Characterization of Klebsiella isolates by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and determination of antimicrobial resistance with VITEK 2 advanced expert system (AES)

Karagöz¹,

Acar²,

Körkoca³

2015

Turk J Med Sci

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Background/aim: The purpose of the study was to evaluate the performance of the VITEK mass spectrometry (MS) (bioMérieux, France) system for the identification of Klebsiella spp. isolated from different sources. Moreover, while assessing the ability of the VITEK 2 automated expert system (AES) to recognize antimicrobial resistance patterns, the researchers have extended the study to compare VITEK 2 with the routine antimicrobial susceptibility testing method. Materials and methods:This study tested 51 Klebsiella spp. isolates that were isolated from environmental examples and clinical examples. Results of conventional methods and the matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) MS were compared. Then, any differing results were compared against a reference 16S rRNA gene sequence, and when indicated, a recA sequencing analysis was done.Results: VITEK MS correctly identified 100% of the Klebsiella spp. isolates. There were two K. oxytoca isolates incorrectly identified to the species level with conventional methods according to the 16S rRNA gene sequencing analysis. In addition, a VITEK 2 AST-N261 card was used for the detection of extended spectrum beta-lactamases (ESBL). Using the VITEK 2 AES, ESBL positivity was found at the rate of 16.3% whereas this rate was 4.08% using the disk diffusion method. Conclusion:MALDI-TOF MS is a rapid and accurate method for the identification of Klebsiella spp. Moreover, the bioMérieux AES provides a useful laboratory tool for the interpretation of susceptibility results.

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Section: Discussionmentioning

confidence: 57%

Characterization of Klebsiella isolates by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and determination of antimicrobial resistance with VITEK 2 advanced expert system (AES)

Karagöz¹,

Acar²,

Körkoca³

2015

Turk J Med Sci

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“…All selected Klebsiella spp. isolates were confirmed prior to analysis using matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF/ MS) as described elsewhere [8].…”

Section: Bacterial Isolationmentioning

confidence: 99%

“…Moreover, an increase in the isolation of MDR strains, especially those producing ESBLs, has also been described [3]. Infections caused by MDR K. pneumoniae strains have been associated with adverse clinical outcomes, including increased mortality, and prolonged hospitalisation associated with economic costs for the health system [8]. Here we characterise ESBLproducing K. pneumoniae isolated from a rural hospital with limited access to third-generation cephalosporins and broad-spectrum antibiotics in general in Southern Mozambique.…”

Section: Introductionmentioning

confidence: 97%

Characterisation of extended-spectrum β-lactamases among Klebsiella pneumoniae isolates causing bacteraemia and urinary tract infection in Mozambique

Pons

Vubil

Guiral

et al. 2015

Journal of Global Antimicrobial Resistance

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“…Protocols for sample preparation and a database to cover such types of veterinary pathogens are needed, and their spectra should be included in the manufacturer's software. Although it is said to be possible to use MALDI-TOF MS for bacterial strain typing, 4,6,41 we have found that it has relatively low discrimination, which may be improved by using a more polymorphic spectrum.…”

Section: Matrix-assisted Laser Desorption/ionization Mass Spectroscopymentioning

confidence: 81%

Nonculture Molecular Techniques for Diagnosis of Bacterial Disease in Animals

2014

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The past decade has seen remarkable technical advances in infectious disease diagnosis, and the pace of innovation is likely to continue. Many of these techniques are well suited to pathogen identification directly from pathologic or clinical samples, which is the focus of this review. Polymerase chain reaction (PCR) and gene sequencing are now routinely performed on frozen or fixed tissues for diagnosis of bacterial infections of animals. These assays are most useful for pathogens that are difficult to culture or identify phenotypically, when propagation poses a biosafety hazard, or when suitable fresh tissue is not available. Multiplex PCR assays, DNA microarrays, in situ hybridization, massive parallel DNA sequencing, microbiome profiling, molecular typing of pathogens, identification of antimicrobial resistance genes, and mass spectrometry are additional emerging technologies for the diagnosis of bacterial infections from pathologic and clinical samples in animals. These technical advances come, however, with 2 caveats. First, in the age of molecular diagnosis, quality control has become more important than ever to identify and control for the presence of inhibitors, cross-contamination, inadequate templates from diagnostic specimens, and other causes of erroneous microbial identifications. Second, the attraction of these technologic advances can obscure the reality that medical diagnoses cannot be made on the basis of molecular testing alone but instead through integrated consideration of clinical, pathologic, and laboratory findings. Proper validation of the method is required. It is critical that veterinary diagnosticians understand not only the value but also the limitations of these technical advances for routine diagnosis of infectious disease.Keywords bacterial infections, mass spectrometry, molecular diagnostic techniques, fluorescent in situ hybridization, quality control, realtime polymerase chain reaction Traditionally, pathogen identification for infectious disease was mostly based on isolation in culture, and this was also critical for fulfilling Koch's postulates. 24 Molecular identification methods have been used for infectious disease diagnosis since the 1980s, and ''molecular Koch's postulates'' were applied to bacterial pathogenicity at the gene rather than the wholeorganism level. 21,22 Molecular guidelines for establishing microbial disease causation were described as a reconsideration of Koch's postulates. 24 Among the nonculture methods, polymerase chain reaction (PCR) and DNA sequencing have been widely used to identify infectious pathogens. Recently, massive parallel DNA sequencing has rapidly developed for the identification of both described and new viral pathogens. The technology will likely revolutionize bacterial disease diagnosis in the near future. Not only have many nonculturable or difficult-to-culture bacterial pathogens become detectable, but many readily isolated bacterial infections can now be identified with nonculture methods. Molecular identification methods ca...

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Biotyping of Multidrug-Resistant Klebsiella pneumoniae Clinical Isolates from France and Algeria Using MALDI-TOF MS

Cited by 76 publications

References 37 publications

Characterization of Klebsiella isolates by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and determination of antimicrobial resistance with VITEK 2 advanced expert system (AES)

Characterization of Klebsiella isolates by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and determination of antimicrobial resistance with VITEK 2 advanced expert system (AES)

Characterisation of extended-spectrum β-lactamases among Klebsiella pneumoniae isolates causing bacteraemia and urinary tract infection in Mozambique

Nonculture Molecular Techniques for Diagnosis of Bacterial Disease in Animals

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