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https://hdl.handle.net/1822/44751
Título: | Discriminating between typical and atypical cystic fibrosis-related bacterial species by multiplex PNA-FISH |
Autor(es): | Lopes, Susana P. Carvalho, Daniel T. Pereira, Maria Olívia Azevedo, Nuno F. |
Palavras-chave: | cystic fibrosis microbiome Fluorescence in situ hybridization Inquilinus limosus PNA probe Pseudomonas aeruginosa |
Data: | Fev-2017 |
Editora: | Wiley-Blackwell |
Revista: | Biotechnology and Bioengineering |
Citação: | Lopes, 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 |
Resumo(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. |
Tipo: | Artigo |
URI: | https://hdl.handle.net/1822/44751 |
DOI: | 10.1002/bit.26085 |
ISSN: | 0006-3592 |
e-ISSN: | 1097-0290 |
Versão da editora: | http://onlinelibrary.wiley.com/journal/10.1002/(ISSN)1097-0290 |
Arbitragem científica: | yes |
Acesso: | Acesso aberto |
Aparece nas coleções: | CEB - Publicações em Revistas/Séries Internacionais / Publications in International Journals/Series |
Ficheiros deste registo:
Ficheiro | Descrição | Tamanho | Formato | |
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document_41156_1.pdf | 2,59 MB | Adobe PDF | Ver/Abrir |