Aptasensor for the detection of Moraxella catarrhalis adhesin UspA2
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/1822/82483 |
Resumo: | Innovative point-of-care (PoC) diagnostic platforms are desirable to surpass the deficiencies of conventional laboratory diagnostic methods for bacterial infections and to tackle the growing antimicrobial resistance crisis. In this study, a workflow was implemented, comprising the identification of new aptamers with high affinity for the ubiquitous surface protein A2 (UspA2) of the bacterial pathogen Moraxella catarrhalis and the development of an electrochemical biosensor functionalized with the best-performing aptamer as a bioreceptor to detect UspA2. After cell-systematic evolution of ligands by exponential enrichment (cell-SELEX) was performed, next-generation sequencing was used to sequence the final aptamer pool. The most frequent aptamer sequences were further evaluated using bioinformatic tools. The two most promising aptamer candidates, Apt1 and Apt1_RC (Apt1 reverse complement), had Kd values of 214.4 and 3.4 nM, respectively. Finally, a simple and label-free electrochemical biosensor was functionalized with Apt1_RC. The aptasensor surface modifications were confirmed by impedance spectroscopy and cyclic voltammetry. The ability to detect UspA2 was evaluated by square wave voltammetry, exhibiting a linear detection range of 4.0 × 104–7.0 × 107 CFU mL−1, a square correlation coefficient superior to 0.99 and a limit of detection of 4.0 × 104 CFU mL−1 at pH 5.0. The workflow described has the potential to be part of a sensitive PoC diagnostic platform to detect and quantify M. catarrhalis from biological samples. |
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Aptasensor for the detection of Moraxella catarrhalis adhesin UspA2adhesinsouter membrane proteins (OMPs)ubiquitous surface protein A2 (UspA2)point-of-care (PoC)biosensorelectrochemicalcell-SELEXaptamersdetectionScience & TechnologyInnovative point-of-care (PoC) diagnostic platforms are desirable to surpass the deficiencies of conventional laboratory diagnostic methods for bacterial infections and to tackle the growing antimicrobial resistance crisis. In this study, a workflow was implemented, comprising the identification of new aptamers with high affinity for the ubiquitous surface protein A2 (UspA2) of the bacterial pathogen Moraxella catarrhalis and the development of an electrochemical biosensor functionalized with the best-performing aptamer as a bioreceptor to detect UspA2. After cell-systematic evolution of ligands by exponential enrichment (cell-SELEX) was performed, next-generation sequencing was used to sequence the final aptamer pool. The most frequent aptamer sequences were further evaluated using bioinformatic tools. The two most promising aptamer candidates, Apt1 and Apt1_RC (Apt1 reverse complement), had Kd values of 214.4 and 3.4 nM, respectively. Finally, a simple and label-free electrochemical biosensor was functionalized with Apt1_RC. The aptasensor surface modifications were confirmed by impedance spectroscopy and cyclic voltammetry. The ability to detect UspA2 was evaluated by square wave voltammetry, exhibiting a linear detection range of 4.0 × 104–7.0 × 107 CFU mL−1, a square correlation coefficient superior to 0.99 and a limit of detection of 4.0 × 104 CFU mL−1 at pH 5.0. The workflow described has the potential to be part of a sensitive PoC diagnostic platform to detect and quantify M. catarrhalis from biological samples.The study received financial support from the ViBrANT project, which received funding from the EU Horizon 2020 Research and Innovation Programme under the Marie Sklowdowska Curie, grant agreement no. 765042. In addition, the authors acknowledge the financial support from Fundação para a Ciência e Tecnologia (FCT) under the scope of the strategic funding of UID/BIO/04469/2020 unit and of LABBELS—Associate Laboratory in Biotechnology, Bioengineering and Microelectromechanical Systems, LA/P/0029/2020. D.L. and A.S. received additional funding from the Research Council of Norway (grant 294605, Center for Digital Life). L.D.R.M. acknowledges funding from the FCT through the Scientific Employment Stimulus Program (2021.00221.CEECIND).info:eu-repo/semantics/publishedVersionMDPIUniversidade do MinhoSande, MariaFerreira, DéboraRodrigues, Joana Lúcia Lima CorreiaMelo, Luís Daniel RodriguesSaragliadis, AthanasiosLinke, DirkMoreira, FelisminaSales, M. G. F.Rodrigues, L. R.2023-01-312023-01-31T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttps://hdl.handle.net/1822/82483engSande, M.G.; Ferreira, D.; Rodrigues, J.L.; Melo, L.D.R.; Saragliadis, A.; Linke, D.; Moreira, F.T.C.; Sales, M.G.F.; Rodrigues, L.R. Aptasensor for the Detection of Moraxella catarrhalis Adhesin UspA2. Bioengineering 2023, 10, 178. https://doi.org/10.3390/bioengineering100201782306-535410.3390/bioengineering10020178https://www.mdpi.com/2306-5354/10/2/178info: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:RCAAP2023-07-21T12:14:27Zoai:repositorium.sdum.uminho.pt:1822/82483Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T19:06:46.777221Repositório 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 |
Aptasensor for the detection of Moraxella catarrhalis adhesin UspA2 |
title |
Aptasensor for the detection of Moraxella catarrhalis adhesin UspA2 |
spellingShingle |
Aptasensor for the detection of Moraxella catarrhalis adhesin UspA2 Sande, Maria adhesins outer membrane proteins (OMPs) ubiquitous surface protein A2 (UspA2) point-of-care (PoC) biosensor electrochemical cell-SELEX aptamers detection Science & Technology |
title_short |
Aptasensor for the detection of Moraxella catarrhalis adhesin UspA2 |
title_full |
Aptasensor for the detection of Moraxella catarrhalis adhesin UspA2 |
title_fullStr |
Aptasensor for the detection of Moraxella catarrhalis adhesin UspA2 |
title_full_unstemmed |
Aptasensor for the detection of Moraxella catarrhalis adhesin UspA2 |
title_sort |
Aptasensor for the detection of Moraxella catarrhalis adhesin UspA2 |
author |
Sande, Maria |
author_facet |
Sande, Maria Ferreira, Débora Rodrigues, Joana Lúcia Lima Correia Melo, Luís Daniel Rodrigues Saragliadis, Athanasios Linke, Dirk Moreira, Felismina Sales, M. G. F. Rodrigues, L. R. |
author_role |
author |
author2 |
Ferreira, Débora Rodrigues, Joana Lúcia Lima Correia Melo, Luís Daniel Rodrigues Saragliadis, Athanasios Linke, Dirk Moreira, Felismina Sales, M. G. F. Rodrigues, L. R. |
author2_role |
author author author author author author author author |
dc.contributor.none.fl_str_mv |
Universidade do Minho |
dc.contributor.author.fl_str_mv |
Sande, Maria Ferreira, Débora Rodrigues, Joana Lúcia Lima Correia Melo, Luís Daniel Rodrigues Saragliadis, Athanasios Linke, Dirk Moreira, Felismina Sales, M. G. F. Rodrigues, L. R. |
dc.subject.por.fl_str_mv |
adhesins outer membrane proteins (OMPs) ubiquitous surface protein A2 (UspA2) point-of-care (PoC) biosensor electrochemical cell-SELEX aptamers detection Science & Technology |
topic |
adhesins outer membrane proteins (OMPs) ubiquitous surface protein A2 (UspA2) point-of-care (PoC) biosensor electrochemical cell-SELEX aptamers detection Science & Technology |
description |
Innovative point-of-care (PoC) diagnostic platforms are desirable to surpass the deficiencies of conventional laboratory diagnostic methods for bacterial infections and to tackle the growing antimicrobial resistance crisis. In this study, a workflow was implemented, comprising the identification of new aptamers with high affinity for the ubiquitous surface protein A2 (UspA2) of the bacterial pathogen Moraxella catarrhalis and the development of an electrochemical biosensor functionalized with the best-performing aptamer as a bioreceptor to detect UspA2. After cell-systematic evolution of ligands by exponential enrichment (cell-SELEX) was performed, next-generation sequencing was used to sequence the final aptamer pool. The most frequent aptamer sequences were further evaluated using bioinformatic tools. The two most promising aptamer candidates, Apt1 and Apt1_RC (Apt1 reverse complement), had Kd values of 214.4 and 3.4 nM, respectively. Finally, a simple and label-free electrochemical biosensor was functionalized with Apt1_RC. The aptasensor surface modifications were confirmed by impedance spectroscopy and cyclic voltammetry. The ability to detect UspA2 was evaluated by square wave voltammetry, exhibiting a linear detection range of 4.0 × 104–7.0 × 107 CFU mL−1, a square correlation coefficient superior to 0.99 and a limit of detection of 4.0 × 104 CFU mL−1 at pH 5.0. The workflow described has the potential to be part of a sensitive PoC diagnostic platform to detect and quantify M. catarrhalis from biological samples. |
publishDate |
2023 |
dc.date.none.fl_str_mv |
2023-01-31 2023-01-31T00:00:00Z |
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/1822/82483 |
url |
https://hdl.handle.net/1822/82483 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Sande, M.G.; Ferreira, D.; Rodrigues, J.L.; Melo, L.D.R.; Saragliadis, A.; Linke, D.; Moreira, F.T.C.; Sales, M.G.F.; Rodrigues, L.R. Aptasensor for the Detection of Moraxella catarrhalis Adhesin UspA2. Bioengineering 2023, 10, 178. https://doi.org/10.3390/bioengineering10020178 2306-5354 10.3390/bioengineering10020178 https://www.mdpi.com/2306-5354/10/2/178 |
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info:eu-repo/semantics/openAccess |
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openAccess |
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application/pdf |
dc.publisher.none.fl_str_mv |
MDPI |
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MDPI |
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