Utilize este identificador para referenciar este registo:
https://hdl.handle.net/1822/82483
Título: | Aptasensor for the detection of Moraxella catarrhalis adhesin UspA2 |
Autor(es): | 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. |
Palavras-chave: | adhesins outer membrane proteins (OMPs) ubiquitous surface protein A2 (UspA2) point-of-care (PoC) biosensor electrochemical cell-SELEX aptamers detection |
Data: | 31-Jan-2023 |
Editora: | MDPI |
Revista: | Bioengineering |
Citação: | 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 |
Resumo(s): | 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. |
Tipo: | Artigo |
URI: | https://hdl.handle.net/1822/82483 |
DOI: | 10.3390/bioengineering10020178 |
e-ISSN: | 2306-5354 |
Versão da editora: | https://www.mdpi.com/2306-5354/10/2/178 |
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_56053_1.pdf | 3,1 MB | Adobe PDF | Ver/Abrir |
Este trabalho está licenciado sob uma Licença Creative Commons