Multilocus Sequence Typing of
            <i>Klebsiella pneumoniae</i>
            Nosocomial Isolates

Diancourt, Laure; Passet, Virginie; Verhoef, J.; Grimont, Patrick A. D.; Brisse, Sylvain

doi:10.1128/jcm.43.8.4178-4182.2005

Cited by 1,172 publications

(886 citation statements)

References 29 publications

(18 reference statements)

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“…The khe gene was amplified using the following primers: forward (5 0 -TGATTGCATTCGCCACTGG-3 0 ) and reverse (5 0 -GGTCAACCCAACGATCCTG-3 0 ) (He et al 2012). The rpoB gene was amplified using the primers VIC3 (5 0 -GGCGAAATGGCWGAGAACCA-3 0 ) and VIC2 (5 0 -GAGTCTTCGAAGTTGTAACC-3 0 ) (Diancourt et al 2005). Each of 50 lL PCR reactions contained 25 lL 29Taq Master Mix (SinoBio, Shanghai, China), 0.4 lM of each primer, and 2 lL genomic DNA.…”

Section: Pcr Protocol and Sequence Determinationmentioning

confidence: 99%

“…Housekeeping genes are very useful in developing multilocus sequence typing (MLST) scheme for discriminating K. pneumoniae, which further provides unambiguous data useful for the epidemiology of K. pneumoniae. For instance, sequences of seven housekeeping genes, i.e., rpoB, gapA, mdh, pgi, phoE, infB and tonB, have been successfully used to develop a MLST scheme for K. pneumoniae (Diancourt et al 2005). On the other hand, the identification of K. pneumoniae strains were generally confirmed in more simple ways, e.g., by rpoB and/or gyrA gene sequences (Brisse and Duijkeren 2005;Brisse and Verhoef 2001;Brisse et al 2004).…”

Section: Introductionmentioning

confidence: 99%

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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: Pcr Protocol and Sequence Determinationmentioning

confidence: 99%

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

View full text Add to dashboard Cite

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“…PCR was performed using primers described previously by Diancourt and coworkers (7). PCR amplicons were treated with ExoSAP-IT (USB Corporation, Cleveland, OH) to remove excess primers and nucleotides.…”

Section: Methodsmentioning

confidence: 99%

Molecular Epidemiology of TwoKlebsiella pneumoniaeMastitis Outbreaks on a Dairy Farm in New York State

et al. 2007

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Klebsiella spp. have become an important cause of clinical mastitis in dairy cows in New York State. We describe the occurrence of two Klebsiella mastitis outbreaks on a single dairy farm. Klebsiella isolates from milk, feces, and environmental sources were compared using random amplified polymorphic DNA (RAPD)-PCR typing. The first mastitis outbreak was caused by a single strain of Klebsiella pneumoniae, RAPD type A, which was detected in milk from eight cows. RAPD type A was also isolated from the rubber liners of milking machine units after milking of infected cows and from bedding in the outbreak pen. Predominance of a single strain could indicate contagious transmission of the organism or exposure of multiple cows to an environmental point source. No new cases with RAPD type A were observed after implementation of intervention measures that targeted the prevention of transmission via the milking machine as well as improvement of environmental hygiene. A second outbreak of Klebsiella mastitis that occurred several weeks later was caused by multiple RAPD types, which rules out contagious transmission and indicates opportunistic infections originating from the environment. The diversity of Klebsiella strains as quantified with Simpson's index of discrimination was significantly higher for isolates from fecal, feed, and water samples than for isolates from milk samples. Several isolates from bedding material that had the phenotypic appearance of Klebsiella spp. were identified as being Raoultella planticola and Raoultella terrigena based on rpoB sequencing.

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“…Additionally, four/eight K. pneumoniae isolates were grouped into the same cluster (Kp1) and two of these isolates also possessed the qnrB2 allele. According to MLST analysis (Diancourt et al 2005), seven/eight K. pneumoniae isolates were typed as ST11 (Table). …”

mentioning

confidence: 99%

Prevalence of plasmid-mediated quinolone resistance determinants among oxyiminocephalosporin-resistant Enterobacteriaceae in Argentina

Cruz

Radice

Sennati

et al. 2013

Mem. Inst. Oswaldo Cruz

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High quinolone resistance rates were observed among oxyiminocephalosporin-resistant enterobacteria. In the present study, we searched for the prevalence of plasmid-mediated quinolone resistance (PMQR) genes within the 55 oxyiminocephalosporin-resistant enterobacteria collected in a previous survey. The main PMQR determinants were aac(6')-Ib-cr and qnrB, which had prevalence rates of 42.4% and 33.3%, respectively. The aac(6')-Ib-cr gene was more frequently found in CTX-M-15-producing isolates, while qnrB was homogeneously distributed among all CTX-M producers.

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Multilocus Sequence Typing of Klebsiella pneumoniae Nosocomial Isolates

Cited by 1,172 publications

References 29 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 Epidemiology of TwoKlebsiella pneumoniaeMastitis Outbreaks on a Dairy Farm in New York State

Prevalence of plasmid-mediated quinolone resistance determinants among oxyiminocephalosporin-resistant Enterobacteriaceae in Argentina

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