A microfluidic platform combined with bacteriophage receptor binding proteins for multiplex detection of Escherichia coli and Pseudomonas aeruginosa in blood
Autor(a) principal: | |
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Data de Publicação: | 2023 |
Outros Autores: | , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | https://hdl.handle.net/1822/80885 |
Resumo: | Bloodstream infections (BSIs) are triggered by the existence of pathogens in blood and are considered a major health burden worldwide, especially when they result in sepsis and septic shock. Common diagnostic methods are time-consuming, present low specificity, or suffer from interference of blood components, which hampers a timely and effective treatment of BSIs. In this work, a novel microfluidic assay was developed combining a bead-based chip and bacteriophage receptor binding proteins (RBPs) as extremely specific and sensitive recognition molecules for the multiplex concentration and detection of Escherichia coli and Pseudomonas aeruginosa, which are highly prevalent bacteria in BSIs. The device comprises a microcolumn in which antibody-functionalized agarose beads were packed allowing the entrapment of the target bacterium from blood, providing its concentration and separation. For bacterial detection, two recombinant RBPs (Gp54 and Gp17) were fused with different fluorescent proteins and used for the identification of P. aeruginosa and E. coli by the measurement of the distinct fluorescent signals obtained. The developed microfluidic-based assay enabled a fast (70min) and highly specific multiplex detection of both pathogens in whole blood, achieving a detection limit of around 103 CFU, without requiring any time-consuming bacterial pre-enrichment step. Furthermore, it provided a quantitative assessment of bacterial loads present in blood. Noteworthy, this miniaturized and inexpensive device presents simple fabrication and operation, showing great potential to be fully automated, demonstrating to be ideal in point-of-care settings. |
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A microfluidic platform combined with bacteriophage receptor binding proteins for multiplex detection of Escherichia coli and Pseudomonas aeruginosa in bloodReceptor binding proteinsmicrofluidic devicesbacterial separation from bloodfluorescence detectionPseudomonas aeruginosaEscherichia coliBacteriophage Receptor binding proteinsScience & TechnologyBloodstream infections (BSIs) are triggered by the existence of pathogens in blood and are considered a major health burden worldwide, especially when they result in sepsis and septic shock. Common diagnostic methods are time-consuming, present low specificity, or suffer from interference of blood components, which hampers a timely and effective treatment of BSIs. In this work, a novel microfluidic assay was developed combining a bead-based chip and bacteriophage receptor binding proteins (RBPs) as extremely specific and sensitive recognition molecules for the multiplex concentration and detection of Escherichia coli and Pseudomonas aeruginosa, which are highly prevalent bacteria in BSIs. The device comprises a microcolumn in which antibody-functionalized agarose beads were packed allowing the entrapment of the target bacterium from blood, providing its concentration and separation. For bacterial detection, two recombinant RBPs (Gp54 and Gp17) were fused with different fluorescent proteins and used for the identification of P. aeruginosa and E. coli by the measurement of the distinct fluorescent signals obtained. The developed microfluidic-based assay enabled a fast (70min) and highly specific multiplex detection of both pathogens in whole blood, achieving a detection limit of around 103 CFU, without requiring any time-consuming bacterial pre-enrichment step. Furthermore, it provided a quantitative assessment of bacterial loads present in blood. Noteworthy, this miniaturized and inexpensive device presents simple fabrication and operation, showing great potential to be fully automated, demonstrating to be ideal in point-of-care settings.The authors acknowledge the funding from the Portuguese Foundation for Science and Technology (FCT) under the scope of the project “Phages‐on‐chip” PTDC/BTM‐SAL/32442/2017 (POCI‐01-0145‐FEDER‐032442) and the strategic funding of the research units CEB (UIDB/04469/2020) and INESC MN (UID/05367/2020) through the pluriannual BASE and PROGRAMATICO financing and BioTecNorte operation (NORTE‐01-0145‐FEDER‐000004) funded by the European Regional Development Fund under the scope of Norte2020 – Programa Operacional Regional do Norte. S.P.C. and C.R.F.C. acknowledge the FCT for the grants SFRH/BD/130098/2017 and PD/BD/135274/2017, respectively.info:eu-repo/semantics/publishedVersionElsevierUniversidade do MinhoCosta, Susana Patrícia ChavesCaneira, Catarina R. F.Chu, VirginiaFreitas, Paulo P.Conde, João P.Carvalho, Carla Alexandra Oliveira Cunha Medeiros2023-022023-02-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttps://hdl.handle.net/1822/80885engCosta, Susana P.; Caneira, Catarina R. F.; Chu, Virginia; Freitas, Paulo P.; Conde, João P.; Carvalho, Carla M., A microfluidic platform combined with bacteriophage receptor binding proteins for multiplex detection of Escherichia coli and Pseudomonas aeruginosa in blood. Sensors and Actuators B-Chemical, 376, Part A(132917), 20230925-400510.1016/j.snb.2022.132917132917https://www.sciencedirect.com/science/article/pii/S092540052201560Xinfo: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:RCAAP2023-07-21T12:41:29Zoai:repositorium.sdum.uminho.pt:1822/80885Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T19:38:28.411836Repositório 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 |
A microfluidic platform combined with bacteriophage receptor binding proteins for multiplex detection of Escherichia coli and Pseudomonas aeruginosa in blood |
title |
A microfluidic platform combined with bacteriophage receptor binding proteins for multiplex detection of Escherichia coli and Pseudomonas aeruginosa in blood |
spellingShingle |
A microfluidic platform combined with bacteriophage receptor binding proteins for multiplex detection of Escherichia coli and Pseudomonas aeruginosa in blood Costa, Susana Patrícia Chaves Receptor binding proteins microfluidic devices bacterial separation from blood fluorescence detection Pseudomonas aeruginosa Escherichia coli Bacteriophage Receptor binding proteins Science & Technology |
title_short |
A microfluidic platform combined with bacteriophage receptor binding proteins for multiplex detection of Escherichia coli and Pseudomonas aeruginosa in blood |
title_full |
A microfluidic platform combined with bacteriophage receptor binding proteins for multiplex detection of Escherichia coli and Pseudomonas aeruginosa in blood |
title_fullStr |
A microfluidic platform combined with bacteriophage receptor binding proteins for multiplex detection of Escherichia coli and Pseudomonas aeruginosa in blood |
title_full_unstemmed |
A microfluidic platform combined with bacteriophage receptor binding proteins for multiplex detection of Escherichia coli and Pseudomonas aeruginosa in blood |
title_sort |
A microfluidic platform combined with bacteriophage receptor binding proteins for multiplex detection of Escherichia coli and Pseudomonas aeruginosa in blood |
author |
Costa, Susana Patrícia Chaves |
author_facet |
Costa, Susana Patrícia Chaves Caneira, Catarina R. F. Chu, Virginia Freitas, Paulo P. Conde, João P. Carvalho, Carla Alexandra Oliveira Cunha Medeiros |
author_role |
author |
author2 |
Caneira, Catarina R. F. Chu, Virginia Freitas, Paulo P. Conde, João P. Carvalho, Carla Alexandra Oliveira Cunha Medeiros |
author2_role |
author author author author author |
dc.contributor.none.fl_str_mv |
Universidade do Minho |
dc.contributor.author.fl_str_mv |
Costa, Susana Patrícia Chaves Caneira, Catarina R. F. Chu, Virginia Freitas, Paulo P. Conde, João P. Carvalho, Carla Alexandra Oliveira Cunha Medeiros |
dc.subject.por.fl_str_mv |
Receptor binding proteins microfluidic devices bacterial separation from blood fluorescence detection Pseudomonas aeruginosa Escherichia coli Bacteriophage Receptor binding proteins Science & Technology |
topic |
Receptor binding proteins microfluidic devices bacterial separation from blood fluorescence detection Pseudomonas aeruginosa Escherichia coli Bacteriophage Receptor binding proteins Science & Technology |
description |
Bloodstream infections (BSIs) are triggered by the existence of pathogens in blood and are considered a major health burden worldwide, especially when they result in sepsis and septic shock. Common diagnostic methods are time-consuming, present low specificity, or suffer from interference of blood components, which hampers a timely and effective treatment of BSIs. In this work, a novel microfluidic assay was developed combining a bead-based chip and bacteriophage receptor binding proteins (RBPs) as extremely specific and sensitive recognition molecules for the multiplex concentration and detection of Escherichia coli and Pseudomonas aeruginosa, which are highly prevalent bacteria in BSIs. The device comprises a microcolumn in which antibody-functionalized agarose beads were packed allowing the entrapment of the target bacterium from blood, providing its concentration and separation. For bacterial detection, two recombinant RBPs (Gp54 and Gp17) were fused with different fluorescent proteins and used for the identification of P. aeruginosa and E. coli by the measurement of the distinct fluorescent signals obtained. The developed microfluidic-based assay enabled a fast (70min) and highly specific multiplex detection of both pathogens in whole blood, achieving a detection limit of around 103 CFU, without requiring any time-consuming bacterial pre-enrichment step. Furthermore, it provided a quantitative assessment of bacterial loads present in blood. Noteworthy, this miniaturized and inexpensive device presents simple fabrication and operation, showing great potential to be fully automated, demonstrating to be ideal in point-of-care settings. |
publishDate |
2023 |
dc.date.none.fl_str_mv |
2023-02 2023-02-01T00:00:00Z |
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 |
https://hdl.handle.net/1822/80885 |
url |
https://hdl.handle.net/1822/80885 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Costa, Susana P.; Caneira, Catarina R. F.; Chu, Virginia; Freitas, Paulo P.; Conde, João P.; Carvalho, Carla M., A microfluidic platform combined with bacteriophage receptor binding proteins for multiplex detection of Escherichia coli and Pseudomonas aeruginosa in blood. Sensors and Actuators B-Chemical, 376, Part A(132917), 2023 0925-4005 10.1016/j.snb.2022.132917 132917 https://www.sciencedirect.com/science/article/pii/S092540052201560X |
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 |
Elsevier |
publisher.none.fl_str_mv |
Elsevier |
dc.source.none.fl_str_mv |
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Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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