Discriminating between typical and atypical cystic fibrosis-related bacterial species by multiplex PNA-FISH
Autor(a) principal: | |
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Data de Publicação: | 2017 |
Outros Autores: | , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/1822/44751 |
Resumo: | 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. |
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Discriminating between typical and atypical cystic fibrosis-related bacterial species by multiplex PNA-FISHcystic fibrosis microbiomeFluorescence in situ hybridizationInquilinus limosusPNA probePseudomonas aeruginosaScience & TechnologyThis 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.Contract grant sponsor: FCT, COMPETE 2020, FEDER, PT2020Contract grant number: UID/BIO/04469/2013Contract grant sponsor: FCT/MEC, FEDER, PT2020Contract grant number: UID/EQU/00511/2013-LEPABEContract grant sponsor: COMPETE 2020Contract grant number: POCI-01-0145-FEDER-006684Contract grant sponsor: FCTContract grant number: RECI/BBB-EBI/0179/2012 (FCOMP-01-0124-FEDER-027462)Contract grant sponsor: QREN, FEDER, ON2Contract grant number: NORTE-07-0124-FEDER-000027Contract grant sponsor: FEDER, ON2, FCT/MECContract grant number: NORTE-07-0124-FEDER-000025Contract grant sponsor: COST-Action TD1004Wiley-BlackwellUniversidade do MinhoLopes, Susana P.Carvalho, Daniel T.Pereira, Maria OlíviaAzevedo, Nuno F.2017-022017-02-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/1822/44751engLopes, 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, 20170006-35921097-029010.1002/bit.2608527571488http://onlinelibrary.wiley.com/journal/10.1002/(ISSN)1097-0290info:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2024-05-11T05:41:55Zoai:repositorium.sdum.uminho.pt:1822/44751Portal AgregadorONGhttps://www.rcaap.pt/oai/openairemluisa.alvim@gmail.comopendoar:71602024-05-11T05:41:55Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
dc.title.none.fl_str_mv |
Discriminating between typical and atypical cystic fibrosis-related bacterial species by multiplex PNA-FISH |
title |
Discriminating between typical and atypical cystic fibrosis-related bacterial species by multiplex PNA-FISH |
spellingShingle |
Discriminating between typical and atypical cystic fibrosis-related bacterial species by multiplex PNA-FISH Lopes, Susana P. cystic fibrosis microbiome Fluorescence in situ hybridization Inquilinus limosus PNA probe Pseudomonas aeruginosa Science & Technology |
title_short |
Discriminating between typical and atypical cystic fibrosis-related bacterial species by multiplex PNA-FISH |
title_full |
Discriminating between typical and atypical cystic fibrosis-related bacterial species by multiplex PNA-FISH |
title_fullStr |
Discriminating between typical and atypical cystic fibrosis-related bacterial species by multiplex PNA-FISH |
title_full_unstemmed |
Discriminating between typical and atypical cystic fibrosis-related bacterial species by multiplex PNA-FISH |
title_sort |
Discriminating between typical and atypical cystic fibrosis-related bacterial species by multiplex PNA-FISH |
author |
Lopes, Susana P. |
author_facet |
Lopes, Susana P. Carvalho, Daniel T. Pereira, Maria Olívia Azevedo, Nuno F. |
author_role |
author |
author2 |
Carvalho, Daniel T. Pereira, Maria Olívia Azevedo, Nuno F. |
author2_role |
author author author |
dc.contributor.none.fl_str_mv |
Universidade do Minho |
dc.contributor.author.fl_str_mv |
Lopes, Susana P. Carvalho, Daniel T. Pereira, Maria Olívia Azevedo, Nuno F. |
dc.subject.por.fl_str_mv |
cystic fibrosis microbiome Fluorescence in situ hybridization Inquilinus limosus PNA probe Pseudomonas aeruginosa Science & Technology |
topic |
cystic fibrosis microbiome Fluorescence in situ hybridization Inquilinus limosus PNA probe Pseudomonas aeruginosa Science & Technology |
description |
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. |
publishDate |
2017 |
dc.date.none.fl_str_mv |
2017-02 2017-02-01T00:00:00Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/1822/44751 |
url |
http://hdl.handle.net/1822/44751 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
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 0006-3592 1097-0290 10.1002/bit.26085 27571488 http://onlinelibrary.wiley.com/journal/10.1002/(ISSN)1097-0290 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Wiley-Blackwell |
publisher.none.fl_str_mv |
Wiley-Blackwell |
dc.source.none.fl_str_mv |
reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação instacron:RCAAP |
instname_str |
Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
instacron_str |
RCAAP |
institution |
RCAAP |
reponame_str |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
collection |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
repository.name.fl_str_mv |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
repository.mail.fl_str_mv |
mluisa.alvim@gmail.com |
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1817544708137680896 |