View More View Less
  • 1 University of Pécs, Pécs, Hungary
  • | 2 University of Pécs, Pécs, Hungary
  • | 3 University of Pécs, Pécs, Hungary
  • | 4 Hungarian Academy of Sciences, Budapest, Hungary
Restricted access

Purchase article

USD  $25.00

1 year subscription (Individual Only)

USD  $784.00

Cross Mark

We investigated the molecular epidemiology of extended spectrum β-lactamase (ESBL) producing Klebsiella pneumoniae isolates derived from the teaching hospitals of University of Pécs, Pécs, Hungary in the time period 2004–2008. Molecular typing, antimicrobial susceptibility testing, detection of common β-lactamase genes (blaCTX-M, blaTEM and blaSHV) and virulence associated traits (hypermucoviscosity, magA, k2a, rmpA, siderophores, type 1 and 3 fimbria, biofilm formation, serum resistance) were performed for 102 isolates. The results showed the presence of three major ciprofloxacin resistant CTX-M-15 producing clones (ST15 n = 69, ST101 n = 10, and ST147 n = 9), of which ST15 was predominant and universally widespread. Considering distribution in time and place, ST101 and ST147 were detected at fewer inpatient units and within a narrower time frame, as compared to ST15. Beside major clones, eleven minor clones were identified, and were shown to harbour the following β-lactamase genes: six clones carried blaCTX-M, four clones harboured blaSHV-5 and one clone possessed both blaCTX-M and ESBL type blaSHV. Among the SHV-5 producing K. pneumoniae clones a novel sequence type was found, namely ST1193, which harboured a unique infB allele. Different virulence factor content and peculiar antimicrobial susceptibility profile were characteristic for each clone. In contrast to major clone isolates, which showed high level resistance to ciprofloxacin, minor clone isolates displayed significantly lower MIC values for ciprofloxacin suggesting a role for fluoroquinolones in the dissemination of the major K. pneumoniae clones. This is the first description of the CTX-M-15 producing K. pneumoniae clone ST101 in Hungary.

  • 1.

    European Centre for Disease Prevention and Control: Antimicrobial resistance surveillance in Europe 2012. Annual Report of the European Antimicrobial Resistance Surveillance Network (EARS-Net) (2013).

    • Search Google Scholar
    • Export Citation
  • 2.

    Coque, T. M., Baquero, F., Canton, R.: Increasing prevalence of ESBL-producing Enterobacteriaceae in Europe. Euro Surveill 13, 573583 (2008).

    • Search Google Scholar
    • Export Citation
  • 3.

    Pragai, Z., Koczian, Z., Nagy, E.: Characterization of the extended-spectrum beta-lactamases and determination of the antibiotic susceptibilities of Klebsiella pneumoniae isolates in Hungary. J Antimicrob Chemother 42, 401403 (1998).

    • Search Google Scholar
    • Export Citation
  • 4.

    Tóth, A., Gacs, M., Márialigeti, K., Cech, G., Füzi, M.: Occurrence and regional distribution of SHV-type extended-spectrum beta-lactamases in Hungary. Eur J Clin Microbiol Infect Dis 24, 284287 (2005).

    • Search Google Scholar
    • Export Citation
  • 5.

    Damjanova, I., Tóth, A., Pászti, J., Jakab, M., Milch, H., Bauernfeind, A., Füzi, M.: Epidemiology of SHV-type beta-lactamase-producing Klebsiella spp. from outbreaks in five geographically distant Hungarian neonatal intensive care units: Widespread dissemination of epidemic R-plasmids. Int J Antimicrob Agents 29, 665671 (2007).

    • Search Google Scholar
    • Export Citation
  • 6.

    Damjanova, I., Tóth, A., Pászti, J., Hajbel-Vékony, G., Jakab, M., Berta, J., Milch, H., Füzi, M.: Expansion and countrywide dissemination of ST11, ST15 and ST147 ciprofloxacin- resistant CTX-M-15-type beta-lactamase-producing Klebsiella pneumoniae epidemic clones in Hungary in 2005 –the new “MRSAs”? J Antimicrob Chemother 62, 978985 (2008).

    • Search Google Scholar
    • Export Citation
  • 7.

    Damjanova, I., Tóth, A., Kenesei, E., Köhalmi, M., Szántai, P., Füzi, M., Pászti, J.: Dissemination of ST274 Klebsiella pneumoniae epidemic clone in newborn and adult hospital settings harbouring SHV-2A or CTX-M-15 type extended spectrum ß-lactamasesproducing known plasmids. Eur J Microbiol Immunol 1, 223227 (2011).

    • Search Google Scholar
    • Export Citation
  • 8.

    Sahly, H., Aucken, H., Benedí, V. J., Forestier, C., Fussing, V., Hansen, D. S., Ofek, I., Podschun, R., Sirot, D., Tomás, J. M., Sandvang, D., Ullmann, U.: Increased serum resistance in Klebsiella pneumoniae strains producing extended-spectrum beta-lactamases. Antimicrob Agents Chemother 48, 34773482 (2004).

    • Search Google Scholar
    • Export Citation
  • 9.

    Sahly, H., Navon-Venezia, S., Roesler, L., Hay, A., Carmeli, Y., Podschun, R., Hennequin, C., Forestier, C., Ofek, I.: Extended-spectrum beta-lactamase production is associated with an increase in cell invasion and expression of fimbrial adhesins in Klebsiella pneumoniae. Antimicrob Agents Chemother 52, 30293034 (2008).

    • Search Google Scholar
    • Export Citation
  • 10.

    Di Martino, P., Livrelli, V., Sirot, D., Joly, B., Darfeuille-Michaud, A.: A new fimbrial antigen harbored by CAZ-5/SHV-4-producing Klebsiella pneumoniae strains involved in nosocomial infections. Infect Immun 64, 22662273 (1996).

    • Search Google Scholar
    • Export Citation
  • 11.

    Shin, J., Ko, K.S.: Comparative study of genotype and virulence in CTX-M-producing and non-extended-spectrum-ß-lactamase-producing Klebsiella pneumoniae isolates. Antimicrob Agents Chemother 58, 24632467 (2014).

    • Search Google Scholar
    • Export Citation
  • 12.

    Di Martino, P., Sirot, D., Joly, B., Rich, C., Darfeuille-Michaud, A.: Relationship between adhesion to intestinal Caco-2 cells and multidrug resistance in Klebsiella pneumoniae clinical isolates. J Clin Microbiol 35, 14991503 (1997).

    • Search Google Scholar
    • Export Citation
  • 13.

    Winn, W. C., Allen, S. D., Janda, W. M., Koneman, E. W., Procop, G. W., Schreckenberger, P. C., Woods, G. L.: Koneman’s Color Atlas and Textbook of Diagnostic Microbiology, 6th Edition. Lippincott Williams & Wilkins, Philadelphia, PA, 2006, pp. 211302.

    • Search Google Scholar
    • Export Citation
  • 14.

    CLSI. Performance Standards for Antimicrobial Susceptibility Testing; Twenty-first Informational Supplement. Document M100-S21. (2009). clsi.org

    • Search Google Scholar
    • Export Citation
  • 15.

    Diancourt, L., Passet, V., Verhoef, J., Grimont, P. A. D., Brisse, S.: Multilocus sequence typing of Klebsiella pneumoniae nosocomial isolates. J Clin Microbiol 43, 41784182 (2005).

    • Search Google Scholar
    • Export Citation
  • 16.

    Edelstein, M., Pimkin, M., Palagin, I., Edelstein, I., Stratchounski, L.: Prevalence and molecular epidemiology of CTX-M extended-spectrum beta-lactamase-producing Escherichia coli and Klebsiella pneumoniae in Russian hospitals. Antimicrob Agents Chemother 47, 37243732 (2003).

    • Search Google Scholar
    • Export Citation
  • 17.

    Bedenic, B., Randegger, C. C., Stobberingh, E., Hächler, H.: Molecular epidemiology of extended-spectrum beta-lactamases from Klebsiella pneumoniae strains isolated in Zagreb, Croatia. Eur J Clin Microbiol Infect Dis 20, 505508 (2001).

    • Search Google Scholar
    • Export Citation
  • 18.

    Wiegand, I., Geiss, H. K., Mack, D., Stürenburg, E., Seifert, H.: Detection of extendedspectrum beta-lactamases among Enterobacteriaceae by use of semiautomated microbiology systems and manual detection procedures. J Clin Microbiol 45, 11671174 (2007).

    • Search Google Scholar
    • Export Citation
  • 19.

    Nüesch-Inderbinen, M. T., Hächler, H., Kayser, F. H.: Detection of genes coding for extended- spectrum SHV beta-lactamases in clinical isolates by a molecular genetic method, and comparison with the E test. Eur J Clin Microbiol Infect Dis 15, 398402 (1996).

    • Search Google Scholar
    • Export Citation
  • 20.

    Jørgensen, R. L., Nielsen, J. B., Friis-Møller, A., Fjeldsøe-Nielsen, H., Schønning, K.: Prevalence and molecular characterization of clinical isolates of Escherichia coli expressing an AmpC phenotype. J Antimicrob Chemother 65, 460464 (2010).

    • Search Google Scholar
    • Export Citation
  • 21.

    The European Committee on Antimicrobial Susceptibility Testing. Breakpoint tables for interpretation of MICs and zone diameters. Version 5.0. (2015). www.eucast.org/clinical_breakpoints

    • Search Google Scholar
    • Export Citation
  • 22.

    Yu, W. L., Fung, C. P., Ko, W. C., Cheng, K. C., Lee, C. C., Chuang, Y. C.: Polymerase chain reaction analysis for detecting capsule serotypes K1 and K2 of Klebsiella pneumoniae causing abscesses of the liver and other sites. J Infect Dis 195, 12351236 (2007).

    • Search Google Scholar
    • Export Citation
  • 23.

    Yeh, K. M., Kurup, A., Siu, L. K., Koh, Y. L., Fung, C. P., Lin, J. C., Chen, T. L., Chang, F. Y., Koh, T. H.: Capsular serotype K1 or K2, rather than magA and rmpA, is a major virulence determinant for Klebsiella pneumoniae liver abscess in Singapore and Taiwan. J Clin Microbiol 45, 466471 (2007).

    • Search Google Scholar
    • Export Citation
  • 24.

    Yu, W. L., Ko, W. C., Cheng, K. C., Lee, H. C., Ke, D. S., Lee, C. C., Fung, C. P., Chuang, Y. C.: Association between rmpA and magA genes and clinical syndromes caused by Klebsiella pneumoniae in Taiwan. Clin Infect Dis 42, 13511358 (2006).

    • Search Google Scholar
    • Export Citation
  • 25.

    Podschun, R., Fischer, A., Ullman, U.: Expression of putative virulence factors by clinical isolates of Klebsiella planticola. J Med Microbiol 49, 115119 (2000).

    • Search Google Scholar
    • Export Citation
  • 26.

    Oelschlaeger, T. A., Zhang, D., Schubert, S., Carniel, E., Rabsch, W., Karch, H., Hacker, J.: The high-pathogenicity island is absent in human pathogens of Salmonella enterica subspecies I but present in isolates of subspecies III and VI. J Bacteriol. 185, 11071111 (2003).

    • Search Google Scholar
    • Export Citation
  • 27.

    Ma, L. C., Fang, C. T., Lee, C. Z., Shun, C. T., Wang, J. T.: Genomic heterogeneity in Klebsiella pneumoniae strains is associated with primary pyogenic liver abscess and metastatic infection. J Infect Dis 192, 117128 (2005).

    • Search Google Scholar
    • Export Citation
  • 28.

    Ofek, I., Beachey, E. H.: Mannose binding and epithelial cell adherence of Escherichia coli. Infect Immun 22, 247254 (1978).

  • 29.

    Schembri, M. A., Blom, J., Krogfelt, K. A., Klemm, P.: Capsule and fimbria interaction in Klebsiella pneumoniae. Infect Immun. 73, 46264633 (2005).

    • Search Google Scholar
    • Export Citation
  • 30.

    Merritt, J. H., Kadouri, D. E., O’Toole, G. A.: Growing and analyzing static biofilms. Curr Protoc Microbiol 00:B:1B.1, 1B.1.1–1B.1.17. (2005).

    • Search Google Scholar
    • Export Citation
  • 31.

    Leclercq, R., Cantón, R., Brown, D. F. J., Giske, C. G., Heisig, P., MacGowan, A. P., Mouton, J. W., Nordmann, P., Rodloff, A. C., Rossolini, G. M., Soussy, C. J., Steinbakk, M., Winstanley, T. G., Kahlmeter, G.: EUCAST expert rules in antimicrobial susceptibility testing. Clin Microbiol Infect 19, 141160 (2013).

    • Search Google Scholar
    • Export Citation
  • 32.

    Donati, V., Feltrin, F., Hendriksen, R. S., Svendsen, C. A., Cordaro, G., García-Fernández, A., Lorenzetti, S., Lorenzetti, R., Battisti, A., Franco, A.: Extended-spectrum-betalactamases, AmpC beta-lactamases and plasmid mediated quinolone resistance in Klebsiella spp. from companion animals in Italy. PLoS One 9, e90564 (2014).

    • Search Google Scholar
    • Export Citation
  • 33.

    Nielsen, J. B., Skov, M. N., Jørgensen, R. L., Heltberg, O., Hansen, D. S., Schønning, K.: Identification of CTX-M15-, SHV-28-producing Klebsiella pneumoniae ST15 as an epidemic clone in the Copenhagen area using a semi-automated Rep-PCR typing assay. Eur J Clin Microbiol Infect Dis 30, 773778 (2011).

    • Search Google Scholar
    • Export Citation
  • 34.

    Rodrigues, C., Machado, E., Ramos, H., Peixe, L., Novais, Â.: Expansion of ESBL-producing Klebsiella pneumoniae in hospitalized patients: A successful story of international clones (ST15, ST147, ST336) and epidemic plasmids (IncR, IncFIIK). Int J Med Microbiol 304, 11001108 (2014).

    • Search Google Scholar
    • Export Citation
  • 35.

    Marcade, G., Brisse, S., Bialek, S., Marcon, E., Leflon-Guibout, V., Passet, V., Moreau, R., Nicolas-Chanoine, M. H.: The emergence of multidrug-resistant Klebsiella pneumoniae of international clones ST13, ST16, ST35, ST48 and ST101 in a teaching hospital in the Paris region. Epidemiol Infect 141, 17051712 (2013).

    • Search Google Scholar
    • Export Citation
  • 36.

    Poulou, A., Voulgari, E., Vrioni, G., Koumaki, V., Xidopoulos, G., Chatzipantazi, V., Markou, F., Tsakris, A.: Outbreak caused by an ertapenem-resistant, CTX-M-15-producing Klebsiella pneumoniae sequence type 101 clone carrying an OmpK36 porin variant. J Clin Microbiol 51, 31763182 (2013).

    • Search Google Scholar
    • Export Citation
  • 37.

    Hrabák, J., Empel, J., Bergerová, T., Fajfrlík, K., Urbásková, P., Kern-Zdanowicz, I., Hryniewicz, W., Gniadkowski, M.: International clones of Klebsiella pneumoniae and Escherichia coli with extended-spectrum beta-lactamases in a Czech hospital. J Clin Microbiol 47, 33533357 (2009).

    • Search Google Scholar
    • Export Citation
  • 38.

    Pitart, C., Solé, M., Roca, I., Fàbrega, A., Vila, J., Marco, F.: First outbreak of a plasmidmediated carbapenem-hydrolyzing OXA-48 beta-lactamase in Klebsiella pneumoniae in Spain. Antimicrob Agents Chemother 55, 43984401 (2011).

    • Search Google Scholar
    • Export Citation
  • 39.

    Lafeuille, E., Decré, D., Mahjoub-Messai, F., Bidet, P., Arlet, G., Bingen, E.: OXA-48 carbapenemase-producing Klebsiella pneumoniae isolated from Libyan patients. Microb Drug Resist 19, 491497 (2013).

    • Search Google Scholar
    • Export Citation
  • 40.

    Potron, A., Poirel, L., Rondinaud, E., Nordmann, P.: Intercontinental spread of OXA-48 beta-lactamase-producing Enterobacteriaceae over an 11-year period, 2001 to 2011. Euro Surveill. 18, ppi:20549 (2013).

    • Search Google Scholar
    • Export Citation
  • 41.

    Frasson, I., Lavezzo, E., Franchin, E., Toppo, S., Barzon, L., Cavallaro, A., Richter, S. N., Palù, G.: Antimicrobial treatment and containment measures for an extremely drug-resistant Klebsiella pneumoniae ST101 isolate carrying pKPN101-IT, a novel fully sequenced bla(KPC-2) plasmid. J Clin Microbiol 50, 37683772 (2012).

    • Search Google Scholar
    • Export Citation
  • 42.

    Oteo, J., Saez, D., Bautista, V., Fernández-Romero, S., Hernández-Molina, J. M., Pérez- Vázquez, M., Aracil, B., Campos, J.: Carbapenemase-producing enterobacteriaceae in Spain in 2012. Antimicrob Agents Chemother 57, 63446347 (2013).

    • Search Google Scholar
    • Export Citation
  • 43.

    Tóth, A., Kocsis, B., Damjanova, I., Kristóf, K., Jánvári, L., Pászti, J., Csercsik, R., Topf, J., Szabó, D., Hamar, P., Nagy, K., Füzi, M.: Fitness cost associated with resistance to fluoroquinolones is diverse across clones of Klebsiella pneumoniae and may select for CTX-M-15 type extended-spectrum ß-lactamase. Eur J Clin Microbiol Infect Dis 33, 837843 (2014).

    • Search Google Scholar
    • Export Citation
  • 44.

    Brisse, S., Fevre, C., Passet, V., Issenhuth-Jeanjean, S., Tournebize, R., Diancourt, L., Grimont, P.: Virulent clones of Klebsiella pneumoniae: Identification and evolutionary scenario based on genomic and phenotypic characterization. PLoS One 4, e4982 (2009).

    • Search Google Scholar
    • Export Citation
  • 45.

    Shin, J., Kim, D. H., Ko, K. S.: Comparison of CTX-M-14- and CTX-M-15-producing Escherichia coli and Klebsiella pneumoniae isolates from patients with bacteremia. J Infect 63, 3947 (2011).

    • Search Google Scholar
    • Export Citation
All Time Past Year Past 30 Days
Abstract Views 217 216 14
Full Text Views 42 23 0
PDF Downloads 32 23 0