Randomly amplified polymorphic DNA typing: a useful tool for rapid epidemiological typing of<i>Klebsiella pneumoniae</i>

Wong, Newton A C S; Linton, Christopher J.; Jalal, Hamid; Millar, Michael

doi:10.1017/s095026880006845x

Cited by 15 publications

(8 citation statements)

References 23 publications

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“…42,2004 MOLECULAR SEROTYPING OF KLEBSIELLA SPECIES ISOLATES 3395 strains belonging to distinct species differed (Table 1 and Fig. 2).…”

Section: Resultsmentioning

confidence: 99%

“…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%

“…42,2004 MOLECULAR SEROTYPING OF KLEBSIELLA SPECIES ISOLATES 3389 In silico restriction analysis and comparative analysis of several restriction enzymes. In order to select restriction enzymes that would generate easily interpretable and informative restriction profiles, the numbers and size distributions of the expected restriction fragments that would be obtained with the two published sequences of the Klebsiella cps cluster were investigated.…”

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confidence: 99%

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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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“…42,2004 MOLECULAR SEROTYPING OF KLEBSIELLA SPECIES ISOLATES 3395 strains belonging to distinct species differed (Table 1 and Fig. 2).…”

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

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

2004

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“…According to Wong et al . [ 28 ], isolates displaying identical RAPD profiles or showing 3 minor band variations were considered the same strain.…”

Section: Methodsmentioning

confidence: 99%

Urinary Tract Infections in Kidney Transplant Patients Due to Escherichia coli and Klebsiella pneumoniae-Producing Extended-Spectrum β-Lactamases: Risk Factors and Molecular Epidemiology

et al. 2015

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Urinary tract infection (UTI) is a common complication after kidney transplantation, often associated to graft loss and increased healthcare costs. Kidney transplant patients (KTPs) are particularly susceptible to infection by Enterobacteriaceae-producing extended-spectrum β-lactamases (ESBLs). A retrospective case-control study was conducted to identify independent risk factors for ESBL-producing Escherichia coli and Klebsiella pneumoniae in non-hospitalized KTPs with UTI. Forty-nine patients suffering from UTI by ESBL-producing bacteria (ESBL-P) as case group and the same number of patients with UTI by ESBL negative (ESBL-N) as control-group were compared. Clinical data, renal function parameters during UTI episodes, UTI recurrence and relapsing rate, as well as risk factors for recurrence, molecular characterization of isolates and the respective antimicrobial susceptibility profile were evaluated. Diabetes mellitus (p <0.007), previous antibiotic prophylaxis (p=0.017) or therapy (p<0.001), previous UTI (p=0.01), relapsing infection (p=0.019) and patients with delayed graft function after transplant (p=0.001) represented risk factors for infection by ESBL positive Enterobacteriaceae in KTPs. Interestingly, the period of time between data of transplantation and data of UTI was shorter in case of ESBL-P case-group (28.8 months) compared with ESBL-N control-group (50.9 months). ESBL-producing bacteria exhibited higher resistance to fluoroquinolones (p=0.002), trimethoprim-sulfamethoxazole (p<0.001) and gentamicin (p<0.001). Molecular analysis showed that bla CTX-M was the most common ESBL encoding gene (65.3%), although in 55.1% of the cases more than one ESBL gene was found. In 29.4% of K. pneumoniae isolates, three bla-genes (bla CTX-M-bla TEM-bla SHV) were simultaneously detected. Low estimated glomerular filtration rate (p=0.009) was found to be risk factor for UTI recurrence. Over 60% of recurrent UTI episodes were caused by genetically similar strains. UTI by ESBL-producing Enterobacteriaceae in KTPs represent an important clinical challenge regarding not only hospitalized patients but also concerning outpatients.

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“…The GenBank accession numbers for the gyrA sequences of strains sb1, sb7, sb9, sb10, sb17, sb18, sb31, sb39, sb55, sb59, sb67, sb69, sb78, sb93, sb113, sb118, sb131, ATCC 13883 S. Brisse and J. Verhoef variation (Mizuta et al, 1983), biotyping (Rennie & Duncan, 1974), protein electrophoretic profiling (Ferragut et al, 1989), multilocus enzyme electrophoresis (Combe et al, 1994), ribotyping (Bingen et al, 1993), randomly amplified polymorphic DNA (RAPD) analysis (Wong et al, 1994), pulsed-field gel electrophoresis (Poh et al, 1993) and diversity of β-lactamase genes (Fournier et al, 1996). The association between genetic variability and virulence and transmissibility of Klebsiella strains is not well understood, but there is clear evidence for differential behaviour of Klebsiella strains.…”

Section: Abbreviationsmentioning

confidence: 99%

Phylogenetic diversity of Klebsiella pneumoniae and Klebsiella oxytoca clinical isolates revealed by randomly amplified polymorphic DNA, gyrA and parC genes sequencing and automated ribotyping.

Brisse¹,

Verhoef²

2001

International Journal of Systematic and Evolutionary Microbiology

240

219

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The infra-specific phylogenetic diversity and genetic structure of both Klebsiella pneumoniae and Klebsiella oxytoca was investigated using a combination of randomly amplified polymorphic DNA (RAPD) analysis, sequencing of gyrA and parC genes, and automated ribotyping. After RAPD analysis with four independent primers of 120 clinical isolates collected from 22 European hospitals in 13 countries, K. pneumoniae isolates fell into three clusters and K. oxytoca isolates fell into two clusters, while Klebsiella planticola isolates formed a sixth cluster. Each cluster was geographically widespread. K. pneumoniae cluster I (KpI) accounted for 80 % of the isolates of this species and included reference strains of the three subspecies K. pneumoniae subsp. pneumoniae, K. pneumoniae subsp. ozaenae and K. pneumoniae subsp. rhinoscleromatis. Clusters KpII and KpIII were equally represented, as were the two K. oxytoca clusters. Individualization of each cluster was fully confirmed by phylogenetic analysis of gyrA and parC gene sequences. In addition, sequence data supported the evolutionary separation of K. pneumoniae from a phylogenetic group including K. oxytoca, Klebsiella terrigena, K. planticola and Klebsiella ornithinolytica. Automated ribotyping using Mlu I appeared suitable for identification of each Klebsiella cluster. The adonitol fermentation test was found to be useful for cluster identification in K. pneumoniae, since it was negative in all strains of clusters KpIII and in some KpII strains, but always positive in cluster KpI. The usefulness of gyrA and parC sequence data for population genetics and cluster identification in bacteria was demonstrated, even for the phylogenetic positioning of quinoloneresistant isolates.

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Randomly amplified polymorphic DNA typing: a useful tool for rapid epidemiological typing ofKlebsiella pneumoniae

Cited by 15 publications

References 23 publications

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

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

Urinary Tract Infections in Kidney Transplant Patients Due to Escherichia coli and Klebsiella pneumoniae-Producing Extended-Spectrum β-Lactamases: Risk Factors and Molecular Epidemiology

Phylogenetic diversity of Klebsiella pneumoniae and Klebsiella oxytoca clinical isolates revealed by randomly amplified polymorphic DNA, gyrA and parC genes sequencing and automated ribotyping.

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