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Electrochemical Aptasensor for the Detection of the Key Virulence Factor YadA of Yersinia enterocolitica

2022Journal article Open access
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dc.contributor.authorSande, Maria Georgina
dc.contributor.authorFerreira, Débora
dc.contributor.authorRodrigues, Joana
dc.contributor.authorMelo, Luís
dc.contributor.authorLinke, Dirk
dc.contributor.authorSilva, Carla J.
dc.contributor.authorMoreira, Felismina
dc.contributor.authorSales, Goreti
dc.contributor.authorRodrigues, Lígia
dc.date.accessioned2023-01-25T16:43:26Z
dc.date.available2023-01-25T16:43:26Z
dc.date.issued2022
dc.description.abstractNew point-of-care (POC) diagnosis of bacterial infections are imperative to overcome the deficiencies of conventional methods, such as culture and molecular methods. In this study, we identified new aptamers that bind to the virulence factor Yersinia adhesin A (YadA) of Yersinia enterocolitica using cell-systematic evolution of ligands by exponential enrichment (cell-SELEX). Escherichia coli expressing YadA on the cell surface was used as a target cell. After eight cycles of selection, the final aptamer pool was sequenced by high throughput sequencing using the Illumina Novaseq platform. The sequencing data, analyzed using the Geneious software, was aligned, filtered and demultiplexed to obtain the key nucleotides possibly involved in the target binding. The most promising aptamer candidate, Apt1, bound specifically to YadA with a dissociation constant (Kd) of 11 nM. Apt1 was used to develop a simple electrochemical biosensor with a two-step, label-free design towards the detection of YadA. The sensor surface modifications and its ability to bind successfully and stably to YadA were confirmed by cyclic voltammetry, impedance spectroscopy and square wave voltammetry. The biosensor enabled the detection of YadA in a linear range between 7.0 × 104 and 7.0 × 107 CFU mL−1 and showed a square correlation coefficient >0.99. The standard deviation and the limit of detection was ~2.5% and 7.0 × 104 CFU mL−1, respectively. Overall, the results suggest that this novel biosensor incorporating Apt1 can potentially be used as a sensitive POC detection system to aid the diagnosis of Y. enterocolitica infections. Furthermore, this simple yet innovative approach could be replicated to select aptamers for other (bacterial) targets and to develop the corresponding biosensors for their detection.pt_PT
dc.description.sponsorshipThis research is affiliated with the VibrANT project that 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 Microelectromechnaical Systems, LA/P/0029/2020.pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.doi10.3390/bios12080614pt_PT
dc.identifier.urihttp://hdl.handle.net/10400.22/21870
dc.language.isoengpt_PT
dc.publisherMDPIpt_PT
dc.relationUID/BIO/04469/2020pt_PT
dc.relationVIral and BacteRial Adhesin Network Training
dc.relation.publisherversionhttps://www.mdpi.com/2079-6374/12/8/614pt_PT
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/4.0/pt_PT
dc.subjectCell-SELEXpt_PT
dc.subjectBiosensorpt_PT
dc.subjectAptamerpt_PT
dc.subjectY. enterocoliticapt_PT
dc.subjectAdhesinpt_PT
dc.subjectYadApt_PT
dc.subjectCyclic voltammetrypt_PT
dc.subjectImpedance spectroscopypt_PT
dc.subjectSquare wave voltammetrypt_PT
dc.titleElectrochemical Aptasensor for the Detection of the Key Virulence Factor YadA of Yersinia enterocoliticapt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.awardTitleVIral and BacteRial Adhesin Network Training
oaire.awardURIinfo:eu-repo/grantAgreement/EC/H2020/765042/EU
oaire.citation.issue8pt_PT
oaire.citation.startPage614pt_PT
oaire.citation.titleBiosensorspt_PT
oaire.citation.volume12pt_PT
oaire.fundingStreamH2020
person.familyNameSande
person.familyNameFerreira
person.familyNameRodrigues
person.familyNameMelo
person.familyNameLinke
person.familyNameMoreira
person.familyNameFerreira Sales
person.familyNameRodrigues
person.givenNameMaria Georgina
person.givenNameDébora
person.givenNameJoana
person.givenNameLuís
person.givenNameDirk
person.givenNameFelismina
person.givenNameMaria Goreti
person.givenNameLigia
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person.identifier.ciencia-idE31F-1630-42BB
person.identifier.ciencia-id8719-B449-CB82
person.identifier.orcid0000-0002-5697-9292
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person.identifier.orcid0000-0003-3150-6752
person.identifier.orcid0000-0003-4237-8952
person.identifier.orcid0000-0001-9936-7336
person.identifier.orcid0000-0001-9265-0630
person.identifier.ridT-1353-2017
person.identifier.ridO-5519-2015
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person.identifier.scopus-author-id27567703100
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person.identifier.scopus-author-id7005174717
person.identifier.scopus-author-id55938875200
project.funder.identifierhttp://doi.org/10.13039/501100008530
project.funder.nameEuropean Commission
rcaap.rightsopenAccesspt_PT
rcaap.typearticlept_PT
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