The DiversiLab system versus pulsed-field gel electrophoresis: Characterisation of extended spectrum β-lactamase producing Escherichia coli and Klebsiella pneumoniae

Brolund, Alma; Hæggman, Sara; Edquist, Petra; Gezelius, Lena; Olsson‐Liljequist, Barbro; Wisell, Karin Tegmark; Giske, Christian G.

doi:10.1016/j.mimet.2010.09.004

Cited by 49 publications

(52 citation statements)

References 18 publications

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“…In a recent publication Carretto et al (2011) found intralaboratory reproducibility to be 98.6 % when the procedure was tested in triplicate, but it was not described how their replicates were performed: three independent DNA samples, three independent rep-PCRs or three different DNA chips. Recent comparisons of DiversiLab rep-PCR typing have been made against multi-locus sequence typing, PFGE and spa-typing (Church et al, 2011;Brolund et al, 2010;Ben-Darif et al, 2010), with the authors concluding that DiversiLab rep-PCR typing is a useful tool for identifying outbreaks. However, using isolates with previously determined Salmonella enterica serotypes, Ben-Darif et al (2010) found that 10 % of their isolates failed to cluster with the correct serotype in the DiversiLab Salmonella serotype library.…”

Section: Discussionmentioning

confidence: 99%

Interlaboratory reproducibility of DiversiLab rep-PCR typing and clustering of Acinetobacter baumannii isolates

Higgins

Hujer

et al. 2012

Journal of Medical Microbiology

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We have investigated the reproducibility of DiversiLab rep-PCR fingerprints between two laboratories with the aim of determining if the fingerprints and clustering are laboratory-specific or portable. One-hundred non-duplicate Acinetobacter baumannii isolates were used in this study. DNA isolation and rep-PCR were each performed separately in two laboratories and rep-PCR patterns generated in laboratory A were compared with those from laboratory B. Twelve A. baumannii isolates processed in laboratory A showed ¢98 % pattern similarity with the corresponding 12 isolates tested in laboratory B and were considered identical. Sixty-four isolates showed 95-97.9 % similarity with their corresponding isolates. Twenty-three isolates showed 90-94 % similarity with the corresponding isolates, while one isolate showed only 87.4 % similarity. However, intra-laboratory clustering was conserved: isolates that clustered in laboratory A also clustered in laboratory B. While clustering was conserved and reproducible at two different laboratories, demonstrating the robustness of rep-PCR, interlaboratory comparison of individual isolate fingerprints showed more variability. This comparison allows conclusions regarding clonality to be reached independent of the laboratory where the analysis is performed.

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Section: Discussionmentioning

confidence: 99%

Interlaboratory reproducibility of DiversiLab rep-PCR typing and clustering of Acinetobacter baumannii isolates

Higgins

Hujer

et al. 2012

Journal of Medical Microbiology

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“…Groups A and B were related, with 84.1% similarity. Previous studies have used PFGE results as low as 80% to 85% to evaluate genetic relationships [24,26]. In this respect, the strains from the non-epidemic period may have been related to the epidemic strains.…”

Section: Discussionmentioning

confidence: 99%

“…Nevertheless, there are some limitations to the use of PFGE, such as its time-consuming nature and requirement for rigorous standardization and experienced personnel to achieve reproducible results that are comparable over time and among locations. Furthermore, there is no consensus on the nomenclature for PFGE patterns and no international database available for comparison [14,24,[27][28][29]. In the present investigation, we used PFGE and rep-PCR for fingerprinting of 15 K. pneumoniae strains and evaluated the results of the two systems.…”

Section: Discussionmentioning

confidence: 99%

“…Additionally, while PFGE showed that strain 9 was not associated with the epidemic, the DL results showed the contrary. Brolund et al [24] indicated that DL can be used to eliminate an unrelated strain in any monitoring study, but the power of this system is not sufficient to discriminate clonally related strains. Confirmation of the results of the DL system by a more powerful system is required.…”

Section: Discussionmentioning

confidence: 99%

“…Homologies of > 97% and 95% to 97% were regarded as the parent clone (indistinguishable) and similar isolates, respectively, according to the manufacturer's recommendations and results of longitudinal studies and comparisons. A similarity value of < 95% was considered to indicate a difference [24].…”

Section: Rep-pcrmentioning

confidence: 99%

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Investigation of clonal relationships of K. pneumoniae isolates from neonatal intensive care units by PFGE and rep-PCR

Köroğlu

Özbek

Demiray

et al. 2015

J Infect Dev Ctries

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Introduction: Clonal relationships of Klebsiella pneumoniae strains obtained during an epidemic and after a one-year post-epidemic (nonepidemic) period in the same neonatal intensive care unit (NICU) using pulsed-field gel electrophoresis (PFGE) and repetitive polymerase chain reaction (rep-PCR) by the DiversiLab (DL) system were investigated, and the results of both molecular techniques were evaluated. Methodology: Fifteen K. pneumoniae strains were included in this study. All identified bacterial strains were confirmed by 16S rDNA sequencing and analyzed by PFGE and the DL system. Results: According to the PFGE results, 15 isolates showed 10 different band profiles. Nine of these 15 isolates were included in one of the formed clusters, and the remaining six isolates were not included in any of them. According to the DL system results, 15 isolates showed two different clusters, with three strains in one cluster and four strains in the other. The remaining strains could not be placed any one of the clusters. PFGE was used as the gold standard based on its strong genetic discriminatory power. The DL system results showed that PFGE missed the relationship of the two epidemic-related strains and demonstrated one epidemic-unrelated strain to be epidemic related. Conclusions: Both systems may easily be used for clonal relationships of K. pneumoniae strains. The DL system was clearly more rapid and convenient than PFGE, but its discriminatory power seemed to be inferior to that of PFGE based on 15 K. pneumoniae strains.

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Emergence of Escherichia coli Sequence Type 131 (ST131) and ST3948 with KPC-2, KPC-3 and KPC-8 carbapenemases from a Long-Term Care and Rehabilitation Facility (LTCRF) in Northern Italy

Piazza

Caltagirone

Bitar

et al. 2015

Advances in Experimental Medicine and Biology

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Aim of the study was to characterize KPC-producing Escherichia coli (KPC-Ec) clinical isolates among a Northern Italy Long-Term Care and Rehabilitation Facility (LTCRF) residents. Thirteen consecutive non repeated MDR E. coli isolates showing ertapenem Minimum Inhibitory Concentrations (MICs) >0.5 mg/L, collected during the period March 2011 - May 2013 from ASP "Redaelli" inpatients, were investigated. The bla KPC/CTX-M/SHV/TEM/OXA genes were identified by PCR and sequencing. KPC-Ec isolates underwent phylotyping, Pulsed-Field Gel Electrophoresis (PFGE), multilocus sequence typing (MLST) and repetitive sequence-based PCR (rep-PCR) profiling. Incompatibility groups analysis and conjugation were also performed. Eleven out of 13 isolates, resulted bla KPC-type positive, were consistently resistant to third generation cephalosporins, fluoroquinolones and trimethoprim-sulphametoxazole (84.6 %), retaining susceptibility to colistin (EUCAST guidelines). At least n = 4/11 of KPC-Ec patients received ≥48 h of meropenem therapy. Sequencing identified 9 bla KPC-2, 1 bla KPC-3 and 1 bla KPC-8 determinants. KPC-Ec plasmids belonged to IncF group (FIIk replicon); conjugation confirmed bla KPC/TEM-1/OXA-9 genes transferability for 10 KPC-Ec. Although three pulsotypes (A, B, C) were identified, all KPC-Ec belonged to phylogenetic group B2. Clone B (B-B5) caused an outbreak of infection involving nine inpatients at five wards. Rep-PCR showed relatedness for seven representative KPC-Ec isolates. Here we report a LTCRF outbreak caused by a ST131-B2 E. coli associated with bla KPC-2 and bla KPC-8 genes, and the emergence of the new ST3948. Elderly people with co-morbidities are at risk for ST131 colonization. KPC-Ec clones local monitoring appears essential both to avoid their spreading among healthcare settings, and to improve therapeutic choices for LTCRF residents.

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The DiversiLab system versus pulsed-field gel electrophoresis: Characterisation of extended spectrum β-lactamase producing Escherichia coli and Klebsiella pneumoniae

Cited by 49 publications

References 18 publications

Interlaboratory reproducibility of DiversiLab rep-PCR typing and clustering of Acinetobacter baumannii isolates

Interlaboratory reproducibility of DiversiLab rep-PCR typing and clustering of Acinetobacter baumannii isolates

Investigation of clonal relationships of K. pneumoniae isolates from neonatal intensive care units by PFGE and rep-PCR

Emergence of Escherichia coli Sequence Type 131 (ST131) and ST3948 with KPC-2, KPC-3 and KPC-8 carbapenemases from a Long-Term Care and Rehabilitation Facility (LTCRF) in Northern Italy

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