Incidence of Burkholderia contaminans at a cystic fibrosis centre with an unusually high representation of Burkholderia cepacia during 15 years of epidemiological surveillance

Coutinho, Carla P.; Barreto, Celeste; Pereira, Luí­sa Álvares; Lito, Luı́s; Melo‐Cristino, José; Sá‐Correia, Isabel

doi:10.1099/jmm.0.000094

Cited by 25 publications

(38 citation statements)

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“…B. contaminans has been isolated worldwide from both environmental and clinical samples (15)(16)(17)(18)(19)(20)(21). B. contaminans has a low prevalence in CF patients worldwide, with remarkable exceptions registered in Argentina (21), Brazil (15,22), Spain (23), and Portugal (17).…”

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“…This fact was attributed to contamination of saline solutions for nasal application (24). Recently, after reexamination of the B. cepacia clinical isolates from the Institute for Bioengineering and Biosciences, Instituto Superior Técnico (iBB/IST) collection, isolates of the recA RFLP type K, formerly classified as B. cepacia species (24, 25), and some additional isolates were classified as B. contaminans (17).The aim of this study was to develop a single nucleotide polymorphism (SNP)-based method for identification and genotyping of B. cepacia and B. contaminans, hereby designated the SNaPBceBcon assay. The methodology implemented allows the clear distinction of these closely related species by testing of a few molecular markers.…”

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confidence: 99%

“…A total of 108 isolates of B. cepacia (n ϭ 83) and B. contaminans (n ϭ 25), obtained at the Portuguese CF treatment center at HSM, in Lisbon, including 35 isolates obtained from contaminated nonsterile saline solutions for nasal application, were used in this study (Table 1) (17,24,25). The isolates had been obtained during the past 2 decades and were characterized molecularly at the iBB/ IST by sequencing analyses of the recA gene (17,(24)(25)(26)(27).…”

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“…The isolates had been obtained during the past 2 decades and were characterized molecularly at the iBB/ IST by sequencing analyses of the recA gene (17,(24)(25)(26)(27). The recA PCR RFLP analysis was also performed by amplification of the entire recA gene (28), amplicons were digested with HaeIII (Amersham Biosciences) restriction endonuclease, and the restriction fragments were separated by electrophoresis in 2% (wt/vol) agarose gels.…”

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SNaPBceBcon : a Practical Tool for Identification and Genotyping of Burkholderia cepacia and Burkholderia contaminans

et al. 2016

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We propose an optimized protocol for an extensive population analysis of Burkholderia cepacia and Burkholderia contaminans. Seven new polymorphisms were added to the recently proposed SNaPBcen assay, and a total of 18 markers ensured the clear identification and distinction of B. cepacia and B. contaminans isolates and high genotypic discrimination (Simpson index of 0.94) compared to those for multilocus sequence typing. The Burkholderia cepacia complex (Bcc) is a very heterogeneous group of 20 bacterial species capable of adapting to an innumerable range of environments (1). The Bcc has emerged as an important opportunistic human pathogen mainly affecting those with serious underlying conditions such as cystic fibrosis (CF) and chronic granulomatous disease (2, 3). A major problem concerning Bcc infections is related to their intrinsic resistance to the antibiotics and biocides most frequently employed clinically, and combined therapy is often needed to treat such infections (4-6). Bcc identification can be challenging even with selective culture media (7,8), a large panel of biochemical tests in routine diagnosis (9), and automated identification systems (e.g., Vitek 2) (10). The recA gene sequence analysis currently offers better species discrimination within the Bcc (11). Multilocus sequence typing (MLST) has also been shown to be excellent for both species identification and strain differentiation (12), but its applicability is limited due to its high cost and time-consuming procedure. An alternative methodology, the SNaPBcen assay, which facilitates large-scale analysis of bacterial isolates, had been proposed for the identification and genotyping of B. cenocepacia (13).In 2009, the novel species B. contaminans and B. lata were proposed for a divergent group formerly classified as B. cepacia recA restriction fragment length polymorphism (RFLP) type K (taxon K) (14). B. contaminans has been isolated worldwide from both environmental and clinical samples (15-21). B. contaminans has a low prevalence in CF patients worldwide, with remarkable exceptions registered in Argentina (21), Brazil (15, 22), Spain (23), and Portugal (17). The epidemiological surveillance of Bcc bacteria in respiratory infections at the major Portuguese CF center of Hospital de Santa Maria (HSM), in Lisbon, identified an exceptionally high representation of B. cepacia species isolates. This fact was attributed to contamination of saline solutions for nasal application (24). Recently, after reexamination of the B. cepacia clinical isolates from the Institute for Bioengineering and Biosciences, Instituto Superior Técnico (iBB/IST) collection, isolates of the recA RFLP type K, formerly classified as B. cepacia species (24, 25), and some additional isolates were classified as B. contaminans (17).The aim of this study was to develop a single nucleotide polymorphism (SNP)-based method for identification and genotyping of B. cepacia and B. contaminans, hereby designated the SNaPBceBcon assay. The methodology implemented allows the clear distinc...

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confidence: 99%

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SNaPBceBcon : a Practical Tool for Identification and Genotyping of Burkholderia cepacia and Burkholderia contaminans

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LipNanoCar Technology – A Versatile and Scalable Technology for the Production of Lipid Nanoparticles

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et al. 2022

Advances in Experimental Medicine and Biology

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Whole Genome Sequence Analysis of Burkholderia contaminans FFH2055 Strain Reveals the Presence of Putative β-Lactamases

et al. 2019

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Burkholderia contaminans is a member of the Burkholderia cepacia complex (Bcc), a pathogen with increasing prevalence among cystic fibrosis (CF) patients and the cause of numerous outbreaks due to the use of contaminated commercial products. The antibiotic resistance determinants, particularly β-lactamases, have been poorly studied in this species. In this work, we explored the whole genome sequence (WGS) of a B. contaminans isolate (FFH 2055) and detected four putative β-lactamase-encoding genes. In general, these genes have more than 93% identity with β-lactamase genes found in other Bcc species. Two β-lactamases, a class A (Pen-like, suggested name PenO) and a class D (OXA-like), were further analyzed and characterized. Amino acid sequence comparison showed that Pen-like has 82% and 67% identity with B. multivorans PenA and B. pseudomallei PenI, respectively, while OXA-like displayed strong homology with class D enzymes within the Bcc, but only 22–44% identity with available structures from the OXA family. PCR reactions designed to study the presence of these two genes revealed a heterogeneous distribution among clinical and industrial B. contaminans isolates. Lastly, blaPenO gene was cloned and expressed into E. coli to investigate the antibiotic resistance profile and confers an extended-spectrum β-lactamase (ESBL) phenotype. These results provide insight into the presence of β-lactamases in B. contaminans, suggesting they play a role in antibiotic resistance of these bacteria.

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Incidence of Burkholderia contaminans at a cystic fibrosis centre with an unusually high representation of Burkholderia cepacia during 15 years of epidemiological surveillance

Cited by 25 publications

References 53 publications

SNaPBceBcon : a Practical Tool for Identification and Genotyping of Burkholderia cepacia and Burkholderia contaminans

SNaPBceBcon : a Practical Tool for Identification and Genotyping of Burkholderia cepacia and Burkholderia contaminans

LipNanoCar Technology – A Versatile and Scalable Technology for the Production of Lipid Nanoparticles

Whole Genome Sequence Analysis of Burkholderia contaminans FFH2055 Strain Reveals the Presence of Putative β-Lactamases

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