Comparison of biotyping, ribotyping, and pulsed-field gel electrophoresis for investigation of a common-source outbreak of Burkholderia pickettii bacteremia

Chetoui, Hafid; Melin, Pierrette; Struelens, Marc; Delhalle, E; Nigo, Masayuki; Ryck, R. De; Mol, Patrick De

doi:10.1128/jcm.35.6.1398-1403.1997

Cited by 34 publications

(10 citation statements)

References 17 publications

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“…Patterns within this cluster differed by a large number of bands; 12 bands if all patterns are considered and 9 bands if only a common pattern is considered. However, the cluster formed at a similarity level (82%) which falls within the strain boundary levels reported for other species (3,23,38,39). If isolates of the same strain may differ by two genetic events (6 band differences) within a 3-month period (41), then within an 11-month period, two further genetic events could have occurred, resulting in up to 12 band differences, as observed in this cluster.…”

Section: Discussionsupporting

confidence: 76%

DNA Banding Pattern Polymorphism in Vancomycin-Resistant Enterococcus faecium and Criteria for Defining Strains

Morrison¹,

Woodford²,

Barrett

et al. 1999

J Clin Microbiol

116

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The degree of DNA banding pattern polymorphism exhibited by vancomycin-resistant Enterococcus faecium (VREM) strains isolated on a renal unit over an 11-month period was investigated. Thirty VREM strains from different patients were analyzed by pulsed-field gel electrophoresis (PFGE; with extended run and optimal pulse times), ribotyping, plasmid profile analysis, biotyping, pyrolysis mass spectrometry, and antibiogram analysis. PFGE resolved 17 banding patterns which formed four distinct clusters at the 82% similarity level. Intercluster band differences ranged from 14 to 31 bands. The strains in one cluster, which contained seven patterns that differed from each other by one to seven bands and from the common pattern by five bands, were confirmed to be a single strain by four of the five other typing methods. The strains in a second cluster with eight patterns, which differed from each other by 1 to 12 bands, contained two subclusters. This subdivision was supported by ribotyping and biotyping. However, it was unclear whether these subclusters represented distinct strains. In one strain, marked polymorphism (patterns that differed from each other by up to four bands) was observed in the ribotype pattern. This study demonstrates the high degree of DNA banding pattern polymorphism found for some strains of VREM and illustrates the complexity involved in defining such strains.

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Section: Discussionsupporting

confidence: 76%

DNA Banding Pattern Polymorphism in Vancomycin-Resistant Enterococcus faecium and Criteria for Defining Strains

Morrison¹,

Woodford²,

Barrett

et al. 1999

J Clin Microbiol

116

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“…The genetic relatedness of isolates was examined by pulsed field gel electrophoresis (PFGE) as previously described (Tenover et al 1995;Chetoui et al 1997). PFGE genotyping of Xba I-digested genomic DNA was performed with a CHEF-DRIII system (Bio-Rad); the images were processed using Gel-Compar II software.…”

Section: Genotyping By Pulsed-field Gel Electrophoresismentioning

confidence: 99%

Oxacillinases and antimicrobial susceptibility of Ralstonia pickettii from pharmaceutical water systems in Croatia

Batarilo

Maravić-Vlahoviček

Bedenić

et al. 2022

Letters in Applied Microbiology

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This study evaluated antibiotic susceptibility and presence of blaOXA22 and blaOXA60 genes in 81 isolates of Ralstonia pickettii obtained from different purified and ultra‐pure water systems in two different geographical areas of Croatia. E‐test and disc diffusion test were performed to determine antibiotic susceptibility. Polymerase chain reaction was applied to detect genes encoding OXA‐22 and OXA‐60 oxacillinases previously identified in R. pickettii. The isolates were genotyped by pulsed‐field gel electrophoresis. The results revealed variable susceptibility/resistance profiles. Our isolates exhibited high susceptibility rates to ceftriaxone, cefotaxime, piperacillin‐tazobactam, ciprofloxacin, imipenem, cefepime and in lesser extent to ceftazidime. High rates of susceptibility were also observed for sulphamethoxazole‐trimethoprim and piperacillin. High resistance rates were noticed for ticarcillin‐clavulanate, aztreonam and meropenem, as well as for all aminoglycosides tested. Modified Hodge test was positive in 51·9% strains, indicating production of carbapenemases. blaOXA22 and blaOXA60 genes were detected in 37·0 and 80·3% strains, respectively. Pulsed‐field gel electrophoresis identified three major clusters containing subclusters. R. pickettii should be taken seriously as a possible cause of nosocomial infections to ensure adequate therapy, to prevent the development of resistant strains and to try to reduce the possibility of R. pickettii surviving in clean and ultra clean water systems.

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“…PFGE patterns of outbreak isolates from patients and washing gloves formed a cluster separate from the unrelated isolates. Outbreak isolates shared 75.7% similarity by Pearson correlation analysis; previous studies used a value of 80% for outbreak grouping (24).…”

Section: Clinical and Major Fatty Acid Characterizationmentioning

confidence: 99%

Phenotypic and Genomic Analyses ofBurkholderia stabilisClinical Contamination, Switzerland

Seth-Smith¹,

Casanova²,

Sommerstein³

et al. 2019

Emerg. Infect. Dis.

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A recent hospital outbreak related to premoistened gloves used to wash patients exposed the difficulties of defining Burkholderia species in clinical settings. The outbreak strain displayed key B. stabilis phenotypes, including the inability to grow at 42°C; we used whole-genome sequencing to confirm the pathogen was B. stabilis. The outbreak strain genome comprises 3 chromosomes and a plasmid, sharing an average nucleotide identity of 98.4% with B. stabilis ATCC27515 BAA-67, but with 13% novel coding sequences. The genome lacks identifiable virulence factors and has no apparent increase in encoded antimicrobial drug resistance, few insertion sequences, and few pseudogenes, suggesting this outbreak was an opportunistic infection by an environmental strain not adapted to human pathogenicity. The diversity among outbreak isolates (22 from patients and 16 from washing gloves) is only 6 single-nucleotide polymorphisms, although the genome remains plastic, with large elements stochastically lost from outbreak isolates. B urkholderia is a diverse genus of gram-negative bacteria, with isolates identified from a variety of environments, and ever more species being identified and classified. Whereas some Burkholderia species are associated with bioremediation potential and antimicrobial and antifungal production, others are animal and human pathogens that generally fall within the B. cepacia complex (Bcc) (1). Burkholderia bacteria have large, flexible, multi-replicon genomes, a large metabolic repertoire, various virulence factors, and inherent resistance to many antimicrobial drugs (2,3). An outbreak of B. stabilis was identified among hospitalized patients across several cantons in Switzerland during 2015-2016 (4). The bacterium caused bloodstream infections, noninvasive infections, and wound contaminations. The source of the infection was traced to contaminated commercially available, premoistened washing gloves used for bedridden patients. After hospitals discontinued use of these gloves, the outbreak resolved. Many instances of Bcc strain contamination of medical devices and solutions have been described (4), including an outbreak in Korea associated with a 0.5% chlorhexidine solution (5). B. stabilis also has been identified in nosocomial infections (6-8). We conducted in-depth characterization of the B. stabilis strain from the Switzerland outbreak by using clinical methods and whole-genome sequencing (WGS). We generated a complete draft genome by combining short-and long-read genomic data and compared it to other outbreak isolates to provide a complete genomic assessment of this strain. We provide a thorough comparative genomic analysis of this outbreak strain. Methods Bacterial Isolate Collection Isolates were collected from 22 patients (labeled 1-22) and 16 contaminated washing gloves (labeled A-P) across Switzerland during the outbreak (4). For comparison, we

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Comparison of biotyping, ribotyping, and pulsed-field gel electrophoresis for investigation of a common-source outbreak of Burkholderia pickettii bacteremia

Cited by 34 publications

References 17 publications

DNA Banding Pattern Polymorphism in Vancomycin-Resistant Enterococcus faecium and Criteria for Defining Strains

DNA Banding Pattern Polymorphism in Vancomycin-Resistant Enterococcus faecium and Criteria for Defining Strains

Oxacillinases and antimicrobial susceptibility of Ralstonia pickettii from pharmaceutical water systems in Croatia

Phenotypic and Genomic Analyses ofBurkholderia stabilisClinical Contamination, Switzerland

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