Discriminating typical and atypical cystic fibrosis-related bacteria by multiplex PNA-FISH.
Biotechnol Bioeng
; 114(2): 355-367, 2017 02.
Article
em En
| MEDLINE
| ID: mdl-27571488
ABSTRACT
This study aims to report the development of peptide nucleic acid (PNA) probes to specifically detect the cystic fibrosis (CF)-associated traditional and atypical species Pseudomonas aeruginosa and Inquilinus limosus, respectively. PNA probes were designed in silico, developed and tested in smears prepared in phosphate-buffer saline (PBS), and in artificial sputum medium (ASM). A multiplex fluorescent in situ hybridization (FISH) approach using the designed probes was further validated in artificially contaminated clinical sputum samples and also applied in polymicrobial 24 h-old biofilms involving P. aeruginosa, I. limosus, and other CF-related bacteria. Both probes showed high predictive and experimental specificities and sensitivities. The multiplex PNA-FISH assay, associated with non-specific staining, was successfully adapted in the clinical samples and in biofilms of CF-related bacteria, allowing differentiating the community members and inferring about microbial-microbial interactions within the consortia. This study revealed the great potential of PNA-FISH as a diagnostic tool to discriminate between classical and less common CF-associated bacteria, being suitable to further describe species-dependent prevention strategies and deliver more effective target control therapeutics. Biotechnol. Bioeng. 2017;114 355-367. © 2016 Wiley Periodicals, Inc.
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Texto completo:
1
Bases de dados:
MEDLINE
Assunto principal:
Bactérias
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Hibridização in Situ Fluorescente
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Biofilmes
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Ácidos Nucleicos Peptídicos
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Fibrose Cística
Tipo de estudo:
Prognostic_studies
Limite:
Humans
Idioma:
En
Revista:
Biotechnol Bioeng
Ano de publicação:
2017
Tipo de documento:
Article
País de afiliação:
Portugal