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      Dissemination of ST274 Klebsiella pneumoniae epidemic clone in newborn and adult hospital settings harbouring SHV-2A or CTX-M-15 type extended spectrum β-lactamases-producing known plasmids

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          Molecular epidemiology and genetic features of an extended-spectrum β-lactamase (ESBL) producing Klebsiella pneumoniae epidemic clone (KP-EC) with elevated ciprofloxacin MIC (minimum inhibitory concentration) values from multiple nosocomial outbreaks and sporadic cases between 2006 and 2008 in Hungary were investigated.

          As a result of continuous monitoring of ESBL-producing KP-ECs, 27 isolates collected from five healthcare facilities were selected for macrorestriction profile analysis by PFGE (pulsed field gel electrophoresis). Of these, 12 strains were isolated from adult inpatients, while 15 strains were from newborns. The MIC values for several antibiotics were determined by agar dilution technique. Molecular typing was further performed by PCR (polymerase chain reaction) and sequencing of several antibiotic resistance genes, plasmid profile analysis, transfer of resistance determinants and multilocus sequence typing (MLST).

          All isolates showed moderate resistance to ciprofloxacin (MICs ranged from 0.5 to 8 mg L −1). PFGE revealed the existence of only one genetic cluster defined as EC IV. PstI digestion of plasmid DNA revealed two highly diverse restriction patterns in “adult” and “newborn” isolates corresponding to plasmids from the Hungarian Epidemic Clone and plasmids isolated from a neonatal nosocomial outbreak in 1998, respectively. Sequence analysis of b-lactamase genes from plasmids of 14 selected isolates detected bla SHV-2a in strains isolated exclusively from newborns and bla CTX-M-15 in strains isolated exclusively from adult inpatients. MLST established that strains of the PFGE cluster belonged to a novel sequence type ST274.

          ESBL-producing K. pneumoniae isolates belonging to the novel sequence type ST274 appeared in the newborn and adult hospital settings in Hungary and acquired SHV-2a or CTX-M-15 type enzymes, respectively. Thus, a new antimicrobial resistance strategy for successful conformation to distinct hospital settings was found.

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          Most cited references 15

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          The worldwide emergence of plasmid-mediated quinolone resistance.

          Fluoroquinolone resistance is emerging in gram-negative pathogens worldwide. The traditional understanding that quinolone resistance is acquired only through mutation and transmitted only vertically does not entirely account for the relative ease with which resistance develops in exquisitely susceptible organisms, or for the very strong association between resistance to quinolones and to other agents. The recent discovery of plasmid-mediated horizontally transferable genes encoding quinolone resistance might shed light on these phenomena. The Qnr proteins, capable of protecting DNA gyrase from quinolones, have homologues in water-dwelling bacteria, and seem to have been in circulation for some time, having achieved global distribution in a variety of plasmid environments and bacterial genera. AAC(6')-Ib-cr, a variant aminoglycoside acetyltransferase capable of modifying ciprofloxacin and reducing its activity, seems to have emerged more recently, but might be even more prevalent than the Qnr proteins. Both mechanisms provide low-level quinolone resistance that facilitates the emergence of higher-level resistance in the presence of quinolones at therapeutic levels. Much remains to be understood about these genes, but their insidious promotion of substantial resistance, their horizontal spread, and their co-selection with other resistance elements indicate that a more cautious approach to quinolone use and a reconsideration of clinical breakpoints are needed.
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            Increasing prevalence of ESBL-producing Enterobacteriaceae in Europe.

            Extended-spectrum beta-lactamases (ESBLs) have been increasingly reported in Europe since their first description in 1983. During the 1990s, they were described mainly as members of the TEM- and SHV-beta-lactamase families in Klebsiella pneumoniae causing nosocomial outbreaks. Nowadays, they are mostly found in Escherichia coli that cause community-acquired infections and with increasing frequency contain CTX-M enzymes. Dissemination of specific clones or clonal groups and epidemic plasmids in community and nosocomial settings has been the main reason for the increase in most of the widespread ESBLs belonging to the TEM (TEM-24, TEM-4, TEM-52), SHV (SHV-5, SHV-12) and CTX-M (CTX-M-9, CTX-M-3, CTX-M-14 or CTX-M-15) families in Europe. Co-selection with other resistances, especially to fluoroquinolones, aminoglycosides and sulfonamides, seems to have contributed to the problem. The emergence of epidemic clones harbouring several beta-lactamases simultaneously (ESBLs, metallo-beta-lactamases or cephamycinases) and of new mechanisms of resistance to fluoroquinolones and aminoglycosides warrants future surveillance studies.
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              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'?

              To investigate the molecular epidemiology of ciprofloxacin-resistant CTX-M-15-producing Klebsiella pneumoniae epidemic clones (ECs) isolated from six nosocomial outbreaks and sporadic cases during 2005 in Hungary. Two hundred and eighty-one extended-spectrum beta-lactamase (ESBL)-producing K. pneumoniae clinical isolates collected from 41 centres were submitted to the National ESBL Reference Laboratory for further investigations. Of the 281 strains, 75 isolates proved to be SHV producers, whereas 6 isolates were ciprofloxacin-susceptible CTX-M-type ESBL producers. One hundred and ninety-six ciprofloxacin-resistant CTX-M-type beta-lactamase-producing isolates collected from 35 centres were subjected to macrorestriction profile analysis. Furthermore, molecular typing was performed by PCR and sequencing of several antibiotic resistance genes, plasmid profile analysis, transfer of resistance determinants and multilocus sequence typing (MLST). PFGE revealed the existence of three genetic clusters defined as ECs, where 129 isolates belonged to the previously described Hungarian EC (HEC), 46 isolates to epidemic clone II (EC II) and 21 isolates to epidemic clone III (EC III), respectively. All isolates harboured plasmids ranging from 2.0 to 230 kb. PstI digestion of plasmid DNA from transconjugants/transformants revealed diverse restriction patterns from distinct ECs. Sequence analysis of beta-lactamase genes from 19 selected isolates detected bla(CTX-M-15) and bla(OXA-1) in strains from all three ECs and bla(TEM-1) in EC III isolates located on large plasmids. ISEcpI associated with CTX-M-15 was detected only on a 50 kb non-conjugative plasmid from EC III. MLST identified three allelic profiles: ST 15 (HEC), ST 11 (EC III) and the novel ST 147 (EC II), which correspond to the PFGE clusters, respectively. In 2005, 97% of all CTX-M-producing K. pneumoniae isolates detected across Hungary were highly ciprofloxacin-resistant CTX-M-15 producers and represented just three stable genetic clones.

                Author and article information

                European Journal of Microbiology and Immunology
                Akadémiai Kiadó, co-published with Springer Science+Business Media B.V., Formerly Kluwer Academic Publishers B.V.
                1 September 2011
                : 1
                : 3
                : 223-227
                [ 1 ] Department of Phage Typing and Molecular Epidemiology, National Center for Epidemiology, Gyáli út 2-6, H-1097, Budapest, Hungary
                [ 2 ] Department of Bacteriology, National Center for Epidemiology, Gyáli út 2-6, H-1097, Budapest, Hungary
                [ 3 ] 1st Department of Pediatrics, Semmelweis University, Bókay János u. 53-54, H-1083, Budapest, Hungary
                [ 4 ] “Jávorszky Ödön” Hospital, Argenti Döme tér 1-3, H-2600, Vác, Hungary
                [ 5 ] Institute of Medical Microbiology, Semmelweis University, Nagyvárad tér 4, H-1089, Budapest, Hungary
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