Genetic profiling of Klebsiella pneumoniae: comparison of pulsed field gel electrophoresis and random amplified polymorphic DNA

Ashayeri-Panah, Mitra; Eftekhar, Fereshteh; Ghamsari, Maryam Mobarak; Parvin, Mahmood; Feizabadi, Mohammad Mehdi

doi:10.1590/s1517-83822013005000055

Cited by 11 publications

(9 citation statements)

References 20 publications

(23 reference statements)

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“…In the same way, several studies between 2008 and 2012 on K. pneumoniae isolates in Iran indicated the genotype diversity among isolates of this bacterium. [ 23 28 34 35 36 37 ] However, there are studies that have reported less diversity and more genotype similarity among isolates. [ 38 39 40 41 ] One explanation could be the fact that the most isolates in the above study were from limited sources in one hospital.…”

Section: Discussionmentioning

confidence: 99%

Dissemination of multidrug-resistant, Class I and II integrons and molecular typing of CTX-M-producing Klebsiella pneumoniae

Akya

Elahi

Chegenelorestani

et al. 2018

Int J App Basic Med Res

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Introduction:Klebsiella pneumoniae (K. pneumoniae) is an important opportunistic pathogen causes serious community and hospital-acquired infections, which is highly resistant to antibiotics. We aimed to determine the frequency of multidrug resistant (MDR) and molecular typing of clinical isolates of K. pneumoniae.Methodology:One hundred isolates of K. pneumoniae were collected from clinical samples in three general hospitals in Kermanshah. The antimicrobial susceptibility and extended-spectrum beta-lactamases (ESBL) production of isolates were determined using disk diffusion and combined disk methods, respectively. The blaCTX-M gene, class I and II integrons were detected using polymerase chain reaction. The blaCTX-M positive isolates were selected for genotyping using pulsed-field gel electrophoresis (PFGE).Results:MDR phenotype was observed in 56% of isolates. The 40% of isolates were ESBL positive and 35 isolates contained blaCTX-M. Class I and II of integrons were detected in 50 (89.2%) and 39 (69.6%) of MDR isolates, respectively. PFGE patterns of K. pneumoniae blaCTX-M positive isolates indicated 19 clusters (X1-19) with different genotype patterns.Conclusions:The study findings highlight the concern of circulating MDR strains of K. pneumoniae with blaCTX-M and class I and II integrons in Kermanshah hospitals. The presence of integrons among isolates may facilitate the spread of new resistance genes in this bacterium. Therefore, surveillance for the spread of MDR strains of this bacterium is recommended in hospitals.

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Section: Discussionmentioning

confidence: 99%

Dissemination of multidrug-resistant, Class I and II integrons and molecular typing of CTX-M-producing Klebsiella pneumoniae

Akya

Elahi

Chegenelorestani

et al. 2018

Int J App Basic Med Res

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“…For RAPD typing, isolates that possessed two or more β-lactamase genes were chosen with the majority of isolates being those recovered in 2015. RAPD-PCR was chosen because it is a rapid and simple method which when optimized has proven competitive with the gold standard of pulse field gel electrophoresis [ 26 , 27 ]. RAPD1, also referred to as Primer 640 by other authors [ 13 , 26 ], proved to be optimal for DNA fingerprinting since it allowed the clear distinction of DNA banding patterns for all of the isolates tested.…”

Section: Discussionmentioning

confidence: 99%

Detection of a CTX-M group 2 beta-lactamase gene in a Klebsiella pneumoniae isolate from a tertiary care hospital, Trinidad and Tobago

Cheddie

Dziva

Akpaka

2017

Ann Clin Microbiol Antimicrob

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BackgroundIdentification of the prevalence and spread of ESBL-mediated antibiotic resistance is essential especially in the hospital setting. It is for this reason, more and more studies are highlighting the importance of complementing phenotypic ESBL-detection techniques with molecular techniques in order to understand the basis and extent of this form of resistance among clinically evolved bacterial populations, especially those belonging to the Enterobacteriaceae family. However, in Trinidad and Tobago and other Caribbean countries, very little is known regarding ESBL detection rates and/or the prevalence of genes conferring ESBL resistance.MethodologySixty-six Klebsiella pneumoniae isolates from clinical specimens phenotypically identified by the Microscan Walkaway-96 System as potential ESBL-producers were analysed in this study. Screening and confirmation of these isolates as ESBL producers was done by the Clinical and Laboratory Standards Institute (CLSI) approved methods. Polymerase chain reaction amplification of beta-lactamase genes bla TEM, bla SHV, bla CTX-M1, bla CTX-M2 and bla AmpC was performed to identify mechanisms of β-lactam resistance.ResultsESBL-producing K. pneumoniae was confirmed in 78.8% (41/52) from isolates collected from a variety of sources during the period, April–July 2015. bla SHV (84.8%) and bla CTX-M (46.9%) were the predominant β-lactamase genes identified. A single K. pneumoniae isolate possessed a bla CTX-M group 2 beta-lactamase gene. RAPD analysis identified a number of epidemiologically related isolates. However, current isolates were unrelated to isolates from previous years.ConclusionThis study revealed that among K. pneumoniae isolates exhibiting extended spectrum β-lactam resistance, there was a high prevalence of bla SHV and bla CTX-M genes. This result highlights the need for a reliable epidemiological apparatus that involves the molecular characterisation of ESBL resistance.Electronic supplementary materialThe online version of this article (doi:10.1186/s12941-017-0209-x) contains supplementary material, which is available to authorized users.

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“…The digested DNA was then separated in a 1% SeaKem Gold agarose (LONZA, Rockland, ME, United States) with pulse time of 6–36 s for 18.5 h at 14°C and constant voltage of 6 V/cm. The patterns were analyzed and compared using BioNumerics software version 6.5 ( Ashayeri-Panah et al, 2013 ). Genotyping for KP1276 was determined using 7 housekeeping genes ( gapA , infB , mdh , pgi , phoE , rpoB , and tonB ), and the MLST method was described previously ( Diancourt et al, 2005 ).…”

Section: Methodsmentioning

confidence: 99%

Characterization of a Novel blaKLUC Variant With Reduced β-Lactam Resistance From an IncA/C Group Plasmid in a Clinical Klebsiella pneumoniae Isolate

Shen

Zhang

et al. 2018

Front. Microbiol.

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Similar to other CTX-M family enzymes, KLUC is a recently identified and emerging determinant of cefotaxime resistance that has been recovered from at least three Enterobacteriaceae species, including Kluyvera cryocrescens, Escherichia coli, and Enterobacter cloacae. Whether this extended-spectrum β-lactamase (ESBL) has been disseminated among commonly isolated Enterobacteriaceae is worthy of further investigation. In this study, we screened 739 nosocomial Enterobacteriaceae isolates (240 Klebsiella pneumoniae and 499 E. coli strains) and found that one K. pneumoniae and four E. coli isolates harbored the blaKLUC gene. Three blaKLUC determinants isolated from E. coli were entirely identical to a blaKLUC-3 gene previously recovered in the same hospital. PFGE of four blaKLUC-harboring E. coli strains showed that prevalence of these determinants was most likely mediated by horizontal gene transfer but not clonal dissemination. However, the variant isolated from K. pneumoniae belonged to a novel member of the KLUC enzyme group. This newly identified enzyme (KLUC-5) has an amino acid substitution compared with previously identified KLUC-1 (G18S) and KLUC-3 (G240D). Antimicrobial susceptibility tests showed that KLUC-5 significantly reduced resistance activity to almost all the selected antimicrobials compared to previously identified KLUC-3. Site-directed mutagenesis showed that blaKLUC-5-D240G and blaKLUC-5-S18G significantly enhanced the MIC against its best substrate. Conjugation and S1-PFGE indicated that blaKLUC-5 was located on a transferable plasmid, which was further decoded by single-molecule, real-time sequencing. Comparative genome analysis showed that its backbone exhibited genetic homology to the IncA/C incompatibility group plasmids. A transposable element, ISEcp1, was detected 256-bp upstream of the blaKLUC-5 gene; this location was inconsistent with the previously identified blaKLUC-1 but congruent with the variants recovered from E. coli in the same hospital. These data provide evidence of the increasingly emerging KLUC group of ESBLs in China.

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Genetic profiling of Klebsiella pneumoniae: comparison of pulsed field gel electrophoresis and random amplified polymorphic DNA

Cited by 11 publications

References 20 publications

Dissemination of multidrug-resistant, Class I and II integrons and molecular typing of CTX-M-producing Klebsiella pneumoniae

Dissemination of multidrug-resistant, Class I and II integrons and molecular typing of CTX-M-producing Klebsiella pneumoniae

Detection of a CTX-M group 2 beta-lactamase gene in a Klebsiella pneumoniae isolate from a tertiary care hospital, Trinidad and Tobago

Characterization of a Novel blaKLUC Variant With Reduced β-Lactam Resistance From an IncA/C Group Plasmid in a Clinical Klebsiella pneumoniae Isolate

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