Development of a PCR probe test for identifying Pseudomonas aeruginosa and Pseudomonas (Burkholderia) cepacia.

O'Callaghan, Elizabeth M.; Tanner, M S; Boulnois, Graham J.

doi:10.1136/jcp.47.3.222

Cited by 37 publications

(25 citation statements)

References 22 publications

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“…O'Callaghan et al [15] also described a PCR assay for I! aeruginosa and B. cepacia identification which used 16s RNA sequence amplification followed by hybridisation with specific oligonucleotide probes.…”

Section: Discussionmentioning

confidence: 99%

PCR identification of Pseudomonas aeruginosa and direct detection in clinical samples from cystic fibrosis patients

Filho¹,

Levi

Bento

et al. 1999

Journal of Medical Microbiology

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This report describes a PCR primer pair that targets the aZgD GDP mannose gene ofPseudomonas aeruginosa and produces a specific 520-bp PCR product useful for R aeruginosa identification. This PCR assay was tested with 182 isolates of I? aeruginosa and 20 isolates of other bacterial species, and demonstrated 100% specificity and sensitivity. The test was also able to detect I? aeruginosa directly in clinical samples such as sputum or throat swabs obtained from cystic fibrosis patients. The combination of this primer with a universal bacterial primer, acting as a control to assess DNA quality in the sample, resulted in a robust PCR method that can be used for rapid l? aeruginosa detection.

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Section: Discussionmentioning

confidence: 99%

PCR identification of Pseudomonas aeruginosa and direct detection in clinical samples from cystic fibrosis patients

Filho¹,

Levi

Bento

et al. 1999

Journal of Medical Microbiology

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“…Clinical and environmental strains were grown overnight on nutrient agar (NA; Difco Laboratories) at 37 and 28°C, respectively. Lysis of the cells of clinical strains was performed by the digestion technique described by O'Callaghan et al (33) in 20 l of lysis buffer (1ϫ Polymed PCR buffer, 0.5% Tween 20, 200 g of proteinase K per ml). Lysis of the cells of rhizosphere B. cepacia strains was performed by the procedure described by Di Cello et al (14).…”

Section: Methodsmentioning

confidence: 99%

Burkholderia cepacia Complex Bacteria from Clinical and Environmental Sources in Italy: Genomovar Status and Distribution of Traits Related to Virulence and Transmissibility

et al. 2002

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Sixty-eight Burkholderia cepacia complex isolates recovered from the sputum of 53 cystic fibrosis patients and 75 isolates collected from the maize rhizosphere were compared to each other to assess their genomovar status as well as some traits related to virulence such as antibiotic susceptibility, proteolytic and hemolytic activities, and transmissibility, in which transmissibility is determined by detection of the esmR and cblA genes. Among the clinical isolates, B. cepacia genomovar III comprised the majority of isolates examined and only a very few isolates were assigned to B. cepacia genomovar I, B. stabilis, and B. pyrrocinia; among the environmental isolates a prevalence of B. cepacia genomovar III and B. ambifaria was observed, whereas few environmental isolates belonging to B. cepacia genomovar I and B. pyrrocinia were found. Antibiotic resistance analysis revealed a certain degree of differentiation between clinical and environmental isolates. Proteolytic activity and onion tissue maceration ability were found to be spread equally among both clinical and environmental isolates, whereas larger percentages of environmental isolates than clinical isolates had hemolytic activity. The esmR gene was found exclusively among isolates belonging to B. cepacia genomovar III, with a marked prevalence in clinical isolates, whereas only one clinical isolate belonging to B. cepacia genomovar III was found to bear the cblA gene. In conclusion, the results of the present study show that the species compositions of the clinical and environmental B. cepacia complex populations examined are quite different and that some of the candidate determinants related to virulence and transmissibility are not confined solely to clinical isolates but are also spread among environmental isolates belonging to different species of the B. cepacia complex.

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“…Several candidate PCR assays aimed at the identification of "B. cepacia" have been described previously (16,51,78,101) but most of these assays were developed before the recognition that the B. cepacia complex consists of several species. In addition, most relied on published DNA sequence data derived from analyses of culture collection strains that, in retrospect, are poorly representative of the total diversity within the B. cepacia complex.…”

Section: Identification Of B Cepacia Complex Organismsmentioning

confidence: 99%

Taxonomy and Identification of the Burkholderia cepacia Complex

et al. 2001

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Development of a PCR probe test for identifying Pseudomonas aeruginosa and Pseudomonas (Burkholderia) cepacia.

Cited by 37 publications

References 22 publications

PCR identification of Pseudomonas aeruginosa and direct detection in clinical samples from cystic fibrosis patients

PCR identification of Pseudomonas aeruginosa and direct detection in clinical samples from cystic fibrosis patients

Burkholderia cepacia Complex Bacteria from Clinical and Environmental Sources in Italy: Genomovar Status and Distribution of Traits Related to Virulence and Transmissibility

Taxonomy and Identification of the Burkholderia cepacia Complex

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