Discrimination of Klebsiella pneumoniae and Klebsiella oxytoca phylogenetic groups and other Klebsiella species by use of amplified fragment length polymorphism

Jonas, Daniel E; Spitzmüller, Bettina; Daschner, Franz; Verhoef, J.; Brisse, Sylvain

doi:10.1016/j.resmic.2003.09.011

Cited by 42 publications

(33 citation statements)

References 32 publications

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“…Furthermore, genetically diverse Enterobacteriaceae species can yield similar phenotypic profiles, resulting in poor discrimination between related species (Garcia-Garrote et al 2000;Hawkey 2006). In this context, genotypic identification may be preferable as this approach is faster and more accurate (e.g., Adékambi et al 2009;Fontana et al 2005;Jonas et al 2004 …”

Section: Discussionmentioning

confidence: 99%

Comparative analyses of phenotypic methods and 16S rRNA, khe, rpoB genes sequencing for identification of clinical isolates of Klebsiella pneumoniae

Guo

Shifei

et al. 2016

Antonie van Leeuwenhoek

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The present work aimed to evaluate 16S rRNA, khe and rpoB gene sequencing for the identification of Klebsiella pneumoniae in comparison with phenotypic methods. Fifteen clinical isolates were examined, which were initially identified as K. pneumoniae subsp. pneumoniae using the automated VITEK 32 system in two hospitals in Enshi City, China. Their identity was further supported by conventional phenotypic methods on the basis of morphological and biochemical characteristics. Using Bayesian phylogenetic analyses and haplotypes network reconstruction, 13 isolates were identified as K.pneumoniae, whereas the other two isolates (K19, K24) were classified as Shigella sp. and Enterobacter sp., respectively. Of the three genes, 16S rRNA and khe gene could discriminate the clinical isolates at the genus level, whereas rpoB could discriminate Klebsiella at the species and even subspecies level. Overall, the gene tree based on rpoB is more compatible with the currently accepted classification of Klebsiella than those based on 16S rRNA and khe genes, showing that rpoB can be a powerful tool for identification of K. pneumoniae isolates. Above all, our study challenges the utility of khe as a species-specific marker for identification of K. pneumoniae.Electronic supplementary material The online version of this article

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Section: Discussionmentioning

confidence: 99%

Comparative analyses of phenotypic methods and 16S rRNA, khe, rpoB genes sequencing for identification of clinical isolates of Klebsiella pneumoniae

Guo

Shifei

et al. 2016

Antonie van Leeuwenhoek

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“…However, due to technical limitations, the practice of serotyping is declining, and molecular methods, such as pulsed-field gel electrophoresis (14,32), amplified fragment polymorphism analysis (16), randomly amplified polymorphic DNA analysis (42), and ribotyping (3,6), are increasingly preferred for use in FIG. 3.…”

Section: Discussionmentioning

confidence: 99%

Molecular Serotyping ofKlebsiellaSpecies Isolates by Restriction of the Amplified Capsular Antigen Gene Cluster

2004

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The objective of the present work was to develop a molecular method that would enable determination of the capsular serotypes of Klebsiella isolates without the use of antiserum. PCR amplification of the capsular antigen gene cluster (cps) was followed by digestion with the restriction enzyme HincII (cps PCR-restriction fragment length polymorphism [RFLP] analysis). The profiles (C patterns) obtained for 224 strains representing the 77 known K serotypes showed 3 to 13 fragments ranging in size from 0.2 to 4.4 kb. A total of 97 distinct C patterns were obtained; 100% of 61 pairs of samples tested twice showed reproducible C patterns. The C patterns were K-type specific; i.e., the C pattern(s) of any K serotype was distinct from the C patterns of all other K serotypes, with the only exceptions being serotypes K22 and K37, which are known to cross-react. For 12 of 17 K types for which at least two strains were included, C-pattern variations were found among strains with the same K serotype. Therefore, cps PCR-RFLP analysis has a higher discriminatory power than classical K serotyping. C-pattern identity was observed among strains with a given K type that were collected many years apart and from distinct sources, indicating C-pattern stability. Only 4.5% of the strains were nontypeable, because of unsuccessful PCR amplification (whereas 8 to 23% are nontypeable by classical K serotyping). Three of four noncapsulated strains analyzed showed recognizable C patterns. The K serotypes of 18 (82%) of 22 recent Klebsiella pneumoniae clinical isolates could be deduced from their C patterns. In conclusion, cps PCR-RFLP analysis allows determination of the K serotype, while it is easier to perform and more discriminatory than classical serotyping.

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“…Comparative studies were carried out to detect the correlation between different capsular serotype, mucoid or non-mucoid phenotype, and the presence of different genetic structures coding for virulence factors or harbored exclusively by virulent strains [10][11][12]. PCR-based subtractive hybridization technique was used to detect genetic sequences carried only by highly-virulent strains of K2 serotype of Klebsiella pneumoniae, and plasmid DNA analysis, amplified fragment length polymorphism (AFLP) pulsed-field gel electrophoresis (PFGE), restriction of the amplified capsular antigen gene cluster and multi locus sequence typing (http://www.mlst.net/) were used as molecular techniques to discriminate between strains [13][14][15].…”

Section: Introductionmentioning

confidence: 99%

Molecular Epidemiology and Virulence Characteristics of Klebsiella pneumoniae Strains Isolated from Hospital-Associated Infections

Damian¹,

Usein²,

Palade³

et al. 2009

TOEPIJ

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Abstract:Purpose: The present study aimed to confirm by classical and molecular laboratory methods hospitalassociated outbreaks due to virulent Klebsiella pneumoniae strains.Methods: Eighty three Klebsiella pneumoniae strains isolated from new-born patients, adults and hospital environment and devices in five hospital units, were analyzed for resistance to antibiotics, including last generation cephalosporins, sensitivity to bacteriophages and pulsed-field gel electrophoresis profiles in order to evaluate the epidemiological relatedness and their clonal spreading. Polymerase chain reaction targeting fur genes and several subtractive sequences (SL 002, SL 003, SL 019, SL 020, SL 021 and SL 025) was used for virulence assessment.Results: More than 50% of strains were resistant to third generation cephalosporins and among them 69% were extended spectrum beta-lactamase producers. Phage typing associated with pulse field gel electrophoresis documented the clonal dispersion of strains. Studying the distribution of virulence sequence, our results reveal that fur gene is present in all strains and among the subtractive sequences the most frequent is SL 020 followed by SL 019. None of the analyzed sequences are present in all clinical isolates and none of the bacterial strains carry all these sequences pointing out the heterogeneity of Klebsiella pneumoniae population. Conclusions:Phage typing method associated with pulse field gel electrophoresis typing and genetic profile for virulence indicated the occurrence of hospital associated-infections produced by Klebsiella pneumoniae strains. Moreover, the results reveal that virulence pattern could be used as a molecular marker in order to define strains which are involved in the process of the development of infectious diseases.

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Discrimination of Klebsiella pneumoniae and Klebsiella oxytoca phylogenetic groups and other Klebsiella species by use of amplified fragment length polymorphism

Cited by 42 publications

References 32 publications

Comparative analyses of phenotypic methods and 16S rRNA, khe, rpoB genes sequencing for identification of clinical isolates of Klebsiella pneumoniae

Comparative analyses of phenotypic methods and 16S rRNA, khe, rpoB genes sequencing for identification of clinical isolates of Klebsiella pneumoniae

Molecular Serotyping ofKlebsiellaSpecies Isolates by Restriction of the Amplified Capsular Antigen Gene Cluster

Molecular Epidemiology and Virulence Characteristics of Klebsiella pneumoniae Strains Isolated from Hospital-Associated Infections

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