Comparason of extraction methods for PCR detection of Burkholderia cepacia complex (BCC) from cystic fibrosis patients

Ergünay, Koray; Yurdakul, Pinar; Şener, Burçin; Özçeli̇k, Uğur; Karabulut, Erdem; Ki̇per, Nural

doi:10.2478/s11536-007-0069-4

Cited by 2 publications

(2 citation statements)

References 21 publications

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“…Plasmid construction and plasmid DNA extraction were carried out according to standard molecular biology techniques (25). Genomic DNA was extracted following the phenol-chloroform method (8). PCR was carried out using a Px2 thermal cycler (Thermo Electron Corporation, Milford, MA).…”

Section: Methodsmentioning

confidence: 99%

Characterization of the Poly-β-1,6- N -Acetylglucosamine Polysaccharide Component of Burkholderia Biofilms

Yakandawala

Gawande

LoVetri

et al. 2011

Appl Environ Microbiol

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We demonstrated the production of poly-␤-1,6-N-acetylglucosamine (PNAG) polysaccharide in the biofilms of Burkholderia multivorans, Burkholderia vietnamiensis, Burkholderia ambifaria, Burkholderia cepacia, and Burkholderia cenocepacia using an immunoblot assay for PNAG. These results were confirmed by further studies, which showed that the PNAG hydrolase, dispersin B, eliminated immunoreactivity of extracts from the species that were tested (B. cenocepacia and B. multivorans). Dispersin B also inhibited biofilm formation and dispersed preformed biofilms of Burkholderia species. These results imply a role for PNAG in the maintenance of Burkholderia biofilm integrity. While PNAG was present in biofilms of all of the wild-type test organisms, a ⌬pgaBC mutant of B. multivorans (Mu5) produced no detectable PNAG, indicating that these genes are needed for Burkholderia PNAG formation. Furthermore, restoration of PNAG production in PNAG negative E. coli TRXWMG⌬C (⌬pgaC) by complementation with B. multivorans pgaBCD confirmed the involvement of these genes in Burkholderia PNAG production. While the confocal scanning laser microscopy of untreated wild-type B. multivorans showed thick, multilayered biofilm, Mu5 and dispersin B-treated wild-type biofilms were thin, poorly developed, and disrupted, confirming the involvement of PNAG in B. multivorans biofilm formation. Thus, PNAG appears to be an important component of Burkholderia biofilms, potentially contributing to its resistance to multiple antibiotics and persistence during chronic infections, including cystic fibrosis-associated infection.

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Section: Methodsmentioning

confidence: 99%

Characterization of the Poly-β-1,6- N -Acetylglucosamine Polysaccharide Component of Burkholderia Biofilms

Yakandawala

Gawande

LoVetri

et al. 2011

Appl Environ Microbiol

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“…The prevalence of the complex was 4.4% of Czech CF patients in year 1996 [2], but during outbreaks in 1997-2003 the number increased to 30% [9,10]. The B. cepacia complex encompasses the most threatening CF pathogens, owing to dramatic increase in symptoms and decline in pulmonary function and causes a potentially fatal necrotizing pneumonia with bacteraemia (so called Cepacia Syndrome) [11].…”

Section: Introductionmentioning

confidence: 99%

Differentiation of pulmonary bacterial pathogens in cystic fibrosis by volatile metabolites emitted by their in vitro cultures: Pseudomonas aeruginosa , Staphylococcus aureus , Stenotrophomonas maltophilia and the Burkholderia cepacia complex

et al. 2016

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As a contribution to the continuing search for breath biomarkers of lung and airways infection in patients with cystic fibrosis, CF, we have analysed the volatile metabolites released in vitro by Pseudomonas aeruginosa and other bacteria involved in respiratory infections in these patients, i.e. those belonging to the Burkholderia cepacia complex, Staphylococcus aureus or Stenotrophomonas maltophilia. These opportunistic pathogens are generally harmless to healthy people but they may cause serious infections in patients with severe underlying disease or impaired immunity such as CF patients. Volatile organic compounds emitted from the cultures of strains belonging to the above-mentioned four taxa were analysed by selected ion flow tube mass spectrometry. In order to minimize the effect of differences in media composition all strains were cultured in three different liquid media. Multivariate statistical analysis reveals that the four taxa can be well discriminated by the differences in the headspace VOC concentration profiles. The compounds that should be targeted in breath as potential biomarkers of airway infection were identified for each of these taxa of CF pathogens.

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Comparason of extraction methods for PCR detection of Burkholderia cepacia complex (BCC) from cystic fibrosis patients

Cited by 2 publications

References 21 publications

Characterization of the Poly-β-1,6- N -Acetylglucosamine Polysaccharide Component of Burkholderia Biofilms

Characterization of the Poly-β-1,6- N -Acetylglucosamine Polysaccharide Component of Burkholderia Biofilms

Differentiation of pulmonary bacterial pathogens in cystic fibrosis by volatile metabolites emitted by their in vitro cultures: Pseudomonas aeruginosa , Staphylococcus aureus , Stenotrophomonas maltophilia and the Burkholderia cepacia complex

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