Discriminating between typical and atypical cystic fibrosis-related bacterial species by multiplex PNA-FISH

Detalhes bibliográficos
Autor(a) principal: Lopes, Susana P.
Data de Publicação: 2017
Outros Autores: Carvalho, Daniel T., Pereira, Maria Olívia, Azevedo, Nuno F.
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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spelling 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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