PCR-Based Identification of Klebsiella pneumoniae subsp. rhinoscleromatis, the Agent of Rhinoscleroma

Fèvre, Cindy; Passet, Virginie; Delétoile, Alexis; Barbe, Valérie; Frangeul, Lionel; Almeida, Ana S.; Sansonetti, Philippe J.; Tournebize, Régis; Brisse, Sylvain

doi:10.1371/journal.pntd.0001052

Cited by 32 publications

(31 citation statements)

References 41 publications

(79 reference statements)

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“…wzi is one of the few genes present in the capsular polysaccharide cluster of all capsular types of K. pneumoniae. The alignment of previously published capsular polysaccharide cluster sequences (17,25,26,41,42) allowed us to identify within the wzi gene a region of high sequence variability flanked by conserved motifs that are suitable for primer design. The 78 K-type reference strains, including strain A1517 (17), were analyzed by PCR using the primers wzi_ for2 and wzi_rev.…”

Section: Resultsmentioning

confidence: 99%

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wzi Gene Sequencing, a Rapid Method for Determination of Capsular Type for Klebsiella Strains

et al. 2013

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f Pathogens of the genus Klebsiella have been classified into distinct capsular (K) types for nearly a century. K typing of Klebsiella species still has important applications in epidemiology and clinical microbiology, but the serological method has strong practical limitations. Our objective was to evaluate the sequencing of wzi, a gene conserved in all capsular types of Klebsiella pneumoniae that codes for an outer membrane protein involved in capsule attachment to the cell surface, as a simple and rapid method for the prediction of K type. The sequencing of a 447-nucleotide region of wzi distinguished the K-type reference strains with only nine exceptions. A reference wzi sequence database was created by the inclusion of multiple strains representing K types associated with high virulence and multidrug resistance. A collection of 119 prospective clinical isolates of K. pneumoniae were then analyzed in parallel by wzi sequencing and classical K typing. Whereas K typing achieved typeability for 81% and discrimination for 94.4% of the isolates, these figures were 98.1% and 98.3%, respectively, for wzi sequencing. The prediction of K type once the wzi allele was known was 94%. wzi sequencing is a rapid and simple method for the determination of the K types of most K. pneumoniae clinical isolates.

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Section: Resultsmentioning

confidence: 99%

“…K-type variation has been linked to the presence or absence of particular genes at the cps locus (17,25,26). The cps locus has a mosaic structure, with a group of six conserved genes (galF, orf2, wzi, wza, wzb, and wzc) at its 5= end.…”

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confidence: 99%

wzi Gene Sequencing, a Rapid Method for Determination of Capsular Type for Klebsiella Strains

et al. 2013

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“…ozaenae and K. pneumoniae subsp. rhinoscleromatis (Fevre et al 2011;Podschun and Ullmann 1998). The two latter subspecies are rarely encountered and are associated with specific diseases (ozena and rhinoscleroma, respectively).…”

Section: Introductionmentioning

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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“…PCR was performed on all isolates to amplify the virulence genes listed in Table 1. Previously described specific primers for K1 and K2 capsular type, rmpA, and wcaG (15), capsular type K3 (16), fimH, uge, wabG, kfu, ureA, allS, and mrkD (17), and aerobactin (iutA) (10) were used in this study.…”

Section: Methodsmentioning

confidence: 99%

Characterization of Pre-Antibiotic Era Klebsiella pneumoniae Isolates with Respect to Antibiotic/Disinfectant Susceptibility and Virulence in Galleria mellonella

Wand

Baker

Benthall

et al. 2015

Antimicrob Agents Chemother

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The EGD Murray collection consists of approximately 500 clinical bacterial isolates, mainly Enterobacteriaceae, isolated from around the world between 1917 and 1949. A number of these "Murray" isolates have subsequently been identified as Klebsiella pneumoniae. Antimicrobial susceptibility testing of these isolates showed that over 30% were resistant to penicillins due to the presence of diverse bla SHV ␤-lactamase genes. Analysis of susceptibility to skin antiseptics and triclosan showed that while the Murray isolates displayed a range of MIC/minimal bactericidal concentration (MBC) values, the mean MIC value was lower than that for more modern K. pneumoniae isolates tested. All Murray isolates contained the cation efflux gene cepA, which is involved in disinfectant resistance, but those that were more susceptible to chlorhexidine were found to have a 9-or 18-bp insertion in this gene. Susceptibility to other disinfectants, e.g., H 2 O 2 , in the Murray isolates was comparable to that in modern K. pneumoniae isolates. The Murray isolates were also less virulent in Galleria and had a different complement of putative virulence factors than the modern isolates, with the exception of an isolate related to the modern lineage CC23. More of the modern isolates (41% compared to 8%) are classified as good/very good biofilm formers, but there was overlap in the two populations. This study demonstrated that a significant proportion of the Murray Klebsiella isolates were resistant to penicillins before their routine use. This collection of pre-antibiotic era isolates may provide significant insights into adaptation in K. pneumoniae in relation to biocide susceptibility.

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PCR-Based Identification of Klebsiella pneumoniae subsp. rhinoscleromatis, the Agent of Rhinoscleroma

Cited by 32 publications

References 41 publications

wzi Gene Sequencing, a Rapid Method for Determination of Capsular Type for Klebsiella Strains

wzi Gene Sequencing, a Rapid Method for Determination of Capsular Type for Klebsiella Strains

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

Characterization of Pre-Antibiotic Era Klebsiella pneumoniae Isolates with Respect to Antibiotic/Disinfectant Susceptibility and Virulence in Galleria mellonella

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