Comparative Population Analysis of Klebsiella pneumoniae Strains with Extended-Spectrum β-Lactamases Colonizing Patients in Rehabilitation Centers in Four Countries

Baraniak, Anna; Izdebski, Radosław; Fiett, Janusz; Sadowy, Ewa; Adler, Amos; Kazma, M.; Salomon, Jérôme; Lawrence, Christine; Rossini, Angelo; Salvia, Antonino; Samsó, Joan Vidal; Fierro, J.; Paul, Mical; Lerman, Yaffa; Malhotra-Kumar, Surbhi; Lammens, Christine; Goossens, Herman; Hryniewicz, Waleria; Brun‐Buisson, C.; Carmeli, Yehuda; Gniadkowski, Marek

doi:10.1128/aac.02571-12

Cited by 49 publications

(40 citation statements)

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“…Novel alleles and STs identified in this study were submitted to the database for coding. CGs and CCs were determined by using the eBURST program (http://eburst.mlst.net) against the MLST database with 1,948 STs (as of 3 July 2015) as described previously (9,13,14). Pulsed-field gel electrophoresis (PFGE) of XbaI-digested genomic DNA was performed on a CHEF-DR III apparatus (Bio-Rad Laboratories, Hercules, CA) according to the instruction manual.…”

Section: Methodsmentioning

confidence: 99%

“…The epidemic drug-resistant clones, called "high-risk" clones (11), may play an important role in the dissemination of antimicrobial resistance. The major international high-risk clones identified by MLST belong to sequence type 11 (ST11), ST15, ST147, and ST258 (11)(12)(13)(14)(15)(16). ST11 and ST258 were included in clonal group (CG) 258 (11,14), while ST15 and ST147 were grouped in CG15 and CG147 (11,13,14), respectively.…”

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confidence: 99%

“…K. pneumoniae is also an important reservoir of antibiotic resistance determinants (1). Several clones of K. pneumoniae with a high prevalence of carbapenem resistance or extended-spectrum ␤-lactamases (ESBLs) have been distributed worldwide (11)(12)(13)(14)(15)(16). The epidemic drug-resistant clones, called "high-risk" clones (11), may play an important role in the dissemination of antimicrobial resistance.…”

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Allocation of Klebsiella pneumoniae Bloodstream Isolates into Four Distinct Groups byompK36Typing in a Taiwanese University Hospital

et al. 2015

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The OmpK36 porin plays a role in carbapenem resistance and may contribute to bacterial virulence in Klebsiella pneumoniae. This study aimed to investigate the characteristics of different groups of K. pneumoniae separated by ompK36 typing. Among 226 nonduplicate K. pneumoniae bloodstream isolates collected at a Taiwanese hospital in 2011, four ompK36 types, designated types A, B, C, and D, were identified by PCR in 61, 28, 100, and 36 isolates, respectively; 1 isolate was untypeable. Statistical analysis showed significantly higher rates of antimicrobial resistance (all tested antibiotics except meropenem), extended-spectrum ␤-lactamases or DHA-1 (47.5% together), Qnr-type quinolone resistance determinants (50.8%), and IncFIIA-type plasmids (49.2%) in group A than in others. Seventeen isolates were identified as belonging to 3 international high-risk clones (4 sequence type 11 [ST11], 10 ST15, and 3 ST147 isolates); all isolates but 1 ST15 isolate were classified in group A. The significant characteristics of group C were hypermucoviscosity (62.0%) and a higher virulence gene content. This group included all serotype K1 (n ‫؍‬ 30), K2 (n ‫؍‬ 25), and K5 (n ‫؍‬ 3) isolates, 6 of 7 K57 isolates, all isolates of major clones associated with pyogenic liver abscesses (29 ST23, 11 ST65, 5 ST86, 7 ST373, and 1 ST375 isolates), and 16 (94.1%) of 17 isolates causing bacteremic liver abscesses. Twelve (42.9%) of the group B isolates were responsible for bacteremic biliary tract infections. Group D was predominant (83.3%) among 12 K20 isolates. This study suggests that most clinical K. pneumoniae isolates can be allocated into four groups with distinct characteristics based on ompK36 types. K lebsiella pneumoniae is an important human opportunistic pathogen that causes a wide variety of community-acquired and nosocomial infections (1-4). The organism is still one of the leading causes of community-acquired pneumonia (3) and has been described as the major cause of pyogenic liver abscesses (PLA) in some Asia-Pacific countries (2, 4). Serotypes K1 and K2 in K. pneumoniae have been linked to invasive infections (2-9). Using multilocus sequence typing (MLST), Brisse et al. (10) identified two virulent K1 clones of K. pneumoniae, clonal complex (CC) 23 and CC82, which were strongly associated with PLA and respiratory infection, respectively. K. pneumoniae is also an important reservoir of antibiotic resistance determinants (1). Several clones of K. pneumoniae with a high prevalence of carbapenem resistance or extended-spectrum ␤-lactamases (ESBLs) have been distributed worldwide (11-16). The epidemic drug-resistant clones, called "high-risk" clones (11), may play an important role in the dissemination of antimicrobial resistance. The major international high-risk clones identified by MLST belong to sequence type 11 (ST11), ST15, ST147,. ST11 and ST258 were included in clonal group (CG) 258 (11,14), while ST15 and ST147 were grouped in CG15 and CG147 (11, 13, 14), respectively.High-level carbapenem resistance in K. pneumoniae ma...

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Section: Methodsmentioning

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Allocation of Klebsiella pneumoniae Bloodstream Isolates into Four Distinct Groups byompK36Typing in a Taiwanese University Hospital

et al. 2015

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“…K. pneumoniae ST336 is a single locus variant of ST17 in the clonal complex 17, whose major clones are ST16, ST17 and ST20 [27][28][29][30]. K. pneumoniae isolates of clonal complex 17 are well recognized contributors to the spread of CTX-M-15 worldwide [31] and ST336 is among the less prevalent yet international clones that have been associated with the spread of various ESBL enzymes, including CTX-M-15, and even carbapenemase resistance [23,32,33].…”

Section: A C C E P T E Dmentioning

confidence: 99%

Long-lasting outbreak due to CTX-M-15-producing Klebsiella pneumoniae ST336 in a rehabilitation ward: report and literature review

Valsdóttir

Jelle

Guðlaugsson

et al. 2017

Journal of Hospital Infection

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“…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)(14)(15)(16)(17)(18)(19)(20)(21). E. coli ATCC 25922 and Klebsiella pneumoniae ATCC 13883 were used as negative controls.…”

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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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Comparative Population Analysis of Klebsiella pneumoniae Strains with Extended-Spectrum β-Lactamases Colonizing Patients in Rehabilitation Centers in Four Countries

Cited by 49 publications

References 47 publications

Allocation of Klebsiella pneumoniae Bloodstream Isolates into Four Distinct Groups byompK36Typing in a Taiwanese University Hospital

Allocation of Klebsiella pneumoniae Bloodstream Isolates into Four Distinct Groups byompK36Typing in a Taiwanese University Hospital

Long-lasting outbreak due to CTX-M-15-producing Klebsiella pneumoniae ST336 in a rehabilitation ward: report and literature review

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

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