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TitleDiscriminating between typical and atypical cystic fibrosis-related bacterial species by multiplex PNA-FISH
Author(s)Lopes, Susana P.
Carvalho, Daniel T.
Pereira, Maria Olívia
Azevedo, Nuno F.
Keywordscystic fibrosis microbiome
Fluorescence in situ hybridization
Inquilinus limosus
PNA probe
Pseudomonas aeruginosa
Issue dateFeb-2017
JournalBiotechnology and Bioengineering
CitationLopes, Susana P.; Carvalho, Daniel T.; Pereira, Maria Olívia; Azevedo, Nuno F., Discriminating between typical and atypical cystic fibrosis-related bacterial species by multiplex PNA-FISH. Biotechnology and Bioengineering, 114(2), 355-376, 2017
Abstract(s)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 and in artificial sputum medium. A multiplex fluorescent in situ hybridization (FISH) approach using the probes was further validated in artificially contaminated clinical sputum samples and also applied to polymicrobial 24h-old biofilms involving the aforementioned and other CF-related bacteria. Both probes showed high predictive and experimental specificities/sensitivities. The multiplex PNA-FISH, 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 emerging CF-associated bacteria, being suitable to further describe species-dependent prevention strategies and deliver more effective target control therapeutics.
Publisher version
AccessOpen access
Appears in Collections:CEB - Publicações em Revistas/Séries Internacionais / Publications in International Journals/Series

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