Molecular and Epidemiological Analysis of Nosocomial Carbapenem-Resistant <i>Klebsiella</i> spp. Using Repetitive Extragenic Palindromic-Polymerase Chain Reaction and Matrix-Assisted Laser Desorption/Ionization-Time of Flight

Treviño, Mercedes; Navarro, Daniel; Barbeito, Gema; García-Riestra, Carlos; Crespo, Carlos; Regueiro, Benito J.

doi:10.1089/mdr.2010.0182

Cited by 17 publications

(19 citation statements)

References 36 publications

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“…In line with recent studies (5,12,17), the results of the present work suggest that MALDI-TOF MS can be used for a rapid and optimal detection of nosocomial A. baumannii outbreaks, as described for other microorganisms (10,11,(13)(14)(15)(16).…”

supporting

confidence: 90%

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Typing of Nosocomial Outbreaks of Acinetobacter baumannii by Use of Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry

et al. 2013

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Acinetobacter baumannii has become a major nosocomial threat, causing infections with high rates of morbidity and mortality (1-4).Acinetobacter baumannii hospital outbreaks have been assessed with various DNA typing methods (5-8). Very few typing methods assess outbreaks in real time. Recently, matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) has been developed for the rapid identification of pathogens (9). There are a growing number of reports showing the utility of MALDI-TOF MS to type different bacterial species for easy and early detection of nosocomial outbreaks (10-16).The aim of this study was to assess the potential utility of MALDI-TOF MS to detect the nosocomial spread of A. baumannii isolates. For this purpose, 35 multidrug-resistant strains of A. baumannii, isolated from colonized or infected patients hospitalized in a general hospital in Italy in 2010, were studied using the repetitive sequence-based PCR (rep-PCR) DiversiLab system (DL; bioMérieux, Marcy l'Etoile, France) and MALDI-TOF MS system (Bruker Daltonics, Bremen, Germany), and the results were compared. After storage at Ϫ70°C, the isolates were thawed out and cultured simultaneously on MacConkey agar.DiversiLab rep-PCR (bioMérieux) was performed according to the manufacturer's instructions (5, 17). Briefly, DNA was extracted from a bacterial culture, amplified using rep-PCR, loaded in LabChip, and run using an Agilent 2100 Bioanalyzer (Agilent Technologies). The results were analyzed by DiversiLab with the Pearson correlation (PC) coefficient to emphasize peak intensities more than peak presence or absence. The strains were considered indistinguishable, similar, or different if the similarity was Ն97.5% without differences in the fingerprinting pattern, Ͼ95% and Ͻ97.5%, or Յ95%, respectively.For MALDI-TOF MS, bacteria were cultured for 24 h at 37°C on MacConkey agar and extracts were prepared as described previously (18, 19). For isolate identification, the row spectra were compared with those in the Biotyper database and a log(score) of Ն2.3 was considered to represent a secure species identification. Spectra were acquired with a microflex LT mass spectrometer (Bruker Daltonics) and recorded in the positive linear mode at a laser frequency of 20 Hz, ion source 1 voltage of 20 kV, ion source 2 voltage of 8.5 kV, and mass range from 2,000 to 20,000 kDa, as described elsewhere (18). Reference spectra of newly created main spectra were added to the original Biotyper database. To evaluate the spectrum variation within each strain and to estimate the mass spectral variance of biological replicates, a principal component analysis (PCA) was performed; two biological replicates from six randomly selected Acinetobacter strains were tested, as described above, and spectra for each selected strain were acquired in the same experiment on the mass spectrometer. Technical mass-spectral variance was also evaluated, analyzing five technical replicates from each Acinetobacter independent culture on the same MALDI t...

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supporting

confidence: 90%

“…Recently, matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) has been developed for the rapid identification of pathogens (9). There are a growing number of reports showing the utility of MALDI-TOF MS to type different bacterial species for easy and early detection of nosocomial outbreaks (10)(11)(12)(13)(14)(15)(16).…”

mentioning

confidence: 99%

Typing of Nosocomial Outbreaks of Acinetobacter baumannii by Use of Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry

et al. 2013

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“…Twenty studies met the inclusion criteria (9)(10)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20)(21)(22)(23)(24)(25)(26)(27)(28). The detailed search process and study selection are depicted in Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Fifteen studies reported on carbapenemase-producing Enterobacteriaceae (9,10,12,14,16,17,(19)(20)(21)(22)(23)(25)(26)(27)(28) and five others on carbapenem-resistant Enterobacteriaceae (11,13,15,18,24). One study included infections due to carbapenem-resistant K. pneumoniae isolates, mainly isolates that produced KPC (11).…”

Section: Resultsmentioning

confidence: 99%

“…In seven studies, the 28-or 30-day mortality was provided (10, 13, 17-19, 25, 27), in one, the 14-day mortality was provided (12), in four, the in-hospital mortality was provided (9,11,15,16), in two, the overall mortality was provided (22,23), in one, the infection-related mortality was provided (28), and in three, the type of mortality provided was not determined (21,24,26).…”

Section: Resultsmentioning

confidence: 99%

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Antibiotic Treatment of Infections Due to Carbapenem-Resistant Enterobacteriaceae: Systematic Evaluation of the Available Evidence

Falagas

Lourida

Poulikakos

et al. 2014

Antimicrob Agents Chemother

322

216

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We sought to evaluate the effectiveness of the antibiotic treatment administered for infections caused by carbapenemase-producing Enterobacteriaceae. The PubMed and Scopus databases were systematically searched. Articles reporting the clinical outcomes of patients infected with carbapenemase-producing Enterobacteriaceae according to the antibiotic treatment administered were eligible. Twenty nonrandomized studies comprising 692 patients who received definitive treatment were included. Almost all studies reported on Klebsiella spp. In 8 studies, the majority of infections were bacteremia, while pneumonia and urinary tract infections were the most common infections in 12 studies. In 10 studies, the majority of patients were critically ill. There are methodological issues, including clinical heterogeneity, that preclude the synthesis of the available evidence using statistical analyses, including meta-analysis. From the descriptive point of view, among patients who received combination treatment, mortality was up to 50% for the tigecycline-gentamicin combination, up to 64% for tigecycline-colistin, and up to 67% for carbapenem-colistin. Among the monotherapy-treated patients, mortality was up to 57% for colistin and up to 80% for tigecycline. Certain regimens were administered to a small number of patients in certain studies. Three studies reporting on 194 critically ill patients with bacteremia showed individually significantly lower mortality in the combination arm than in the monotherapy arm. In the other studies, no significant difference in mortality was recorded between the compared groups. Combination antibiotic treatment may be considered the optimal option for severely ill patients with severe infections. However, well-designed randomized studies of specific patient populations are needed to further clarify this issue.

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Elucidating constraints for differentiation of major human Klebsiella pneumoniae clones using MALDI-TOF MS

Rodrigues

Novais

Sousa

et al. 2016

Eur J Clin Microbiol Infect Dis

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The establishment of matrix-assisted laser desorption time-of-flight mass spectrometry (MALDI-TOF MS) in routine microbial identification boosted many developments towards high-throughput applications, including bacterial typing. However, results are still controversial for different bacterial species. We aim to evaluate the suitability of MALDI-TOF MS for typing clinically relevant multidrug resistant (MDR) Klebsiella pneumoniae subsp. pneumoniae clones using routine conditions and a previously validated chemometric analysis workflow. Mass spectra of 83 K. pneumoniae clinical isolates representing major human MDR clones [11 sequence types (STs), 22 PFGE-types] recovered in Portugal and Spain during outbreaks and non-outbreak situations (2003-2012) were obtained from cell extracts (CE) and intact cells (IC), and analysed with different chemometric tools. We observed a highly consistent peak pattern among isolates from different clones either with CE or IC, suggesting a high degree of conservation of biomolecules analysed (a large part corresponding to ribosomal proteins). Moreover, the low degree of agreement between MALDI-TOF MS and other methods (from 34.9 % to 43.4 % of correct assignments for CE and from 40.8 % to 70.1 % for IC) corroborates the low discriminatory potential of the technique at infraspecies level. Our results suggest a low discriminatory power of MALDI-TOF MS for clinically relevant MDR K. pneumoniae clones and highlight the need of developing tools for high-resolution typing in this species.

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Molecular and Epidemiological Analysis of Nosocomial Carbapenem-Resistant Klebsiella spp. Using Repetitive Extragenic Palindromic-Polymerase Chain Reaction and Matrix-Assisted Laser Desorption/Ionization-Time of Flight

Cited by 17 publications

References 36 publications

Typing of Nosocomial Outbreaks of Acinetobacter baumannii by Use of Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry

Typing of Nosocomial Outbreaks of Acinetobacter baumannii by Use of Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry

Antibiotic Treatment of Infections Due to Carbapenem-Resistant Enterobacteriaceae: Systematic Evaluation of the Available Evidence

Elucidating constraints for differentiation of major human Klebsiella pneumoniae clones using MALDI-TOF MS

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