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Direct PCR-free electrochemical biosensing of plant-food derived nucleic acids in genomic DNA extracts. Application to the determination of the key allergen Sola l 7 in tomato seeds

2019-05-07Journal article Open access
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dc.contributor.authorPereira-Barros, Magda A.
dc.contributor.authorBarroso, M. Fátima
dc.contributor.authorMartín-Pedraza, Laura
dc.contributor.authorVargas, Eva
dc.contributor.authorBenedé, Sara
dc.contributor.authorVillalba, Mayte
dc.contributor.authorRocha, João M.
dc.contributor.authorCampuzano, Susana
dc.contributor.authorPingarrón, José M.
dc.date.accessioned2020-06-25T10:53:05Z
dc.date.available2020-06-25T10:53:05Z
dc.date.issued2019-05-07
dc.description.abstractA novel and disposable electrochemical biosensor for PCR-free and selective detection of Sola l 7, a non-specific lipid transfer protein (nsLTP) found in tomato seeds associated to severe symptoms of tomato-allergic patients, is reported in this work. The methodology involves the formation of DNA/RNA heterohybrids by sandwich hybridization of a specific fragment of the Sola l 7 allergen coding sequence with appropriate RNA probes designed and described for the first time in this work. Labeling was carried out with commercial antibodies specific to the heteroduplexes and secondary antibodies conjugated with HRP onto the surface of magnetic beads. Amperometric transduction was performed upon magnetic capture of the resulting magnetic bioconjugates on screen-printed electrodes using the system H2O2/HQ. A comparison of the sandwich hybridization format with a direct approach as well as between different labeling strategies was performed. The LOD value achieved was 0.2 pM (5 amol in 25 μL). The biosensor was successfully applied to the selective analysis of the targeted Sola l 7 specific region directly in just 100 ng of non-fragmented denatured genomic DNA extracted from tomato seeds.pt_PT
dc.description.sponsorshipThe financial support of the Spanish Ministerio de Economía y Competitividad Research Projects, CTQ2015-64402-C2-1-R, SAF2017-86483-R and the TRANSNANOAVANSENS Program from the Comunidad de Madrid (Grant P2018/NMT-4349) are gratefully acknowledged. M.F.B. is grateful to FCT grant SFRH/BPD/78845/2011,financed by POPH–QREN–Tipologia4.1–Formação Avançada, subsidized by Fundo Social Europeu and Ministério da Ciência, Tecnologiae Ensino Superior. The author M.A.P.B. is grateful to the authors J.M.R., M.F.B., S.C., J.M.P. for the scientific assistance and suggestions shared throughout the supervision of her Master’s project and Master’s thesis at University of Minho. The author M.A.P.B. also acknowledges the Department of Biology (DB) and the Centre of Molecular and Environmental Biology (CBMA) from University of Minho (UM), Braga,Portugal, by providing all the conditions leading to the Master's thesis in“Biophysics and Bionanosystems”. Financial support of M.A.P.B. forthis work was provided by a fellowship within the programme Erasmus+ scholarship/Portugal, which allowed performing the laboratorial work at the University Complutense of Madrid, Madrid, Spain. The author J.M.R. acknowledges CBMA-UM and DB-UM, Portugal, by the conditions provided. J.M.R. is also grateful to the Green Chemistry Laboratory (LAQV)–Research center Chemistry and Technology Network (REQUIMTE), and the Department of Chemistry and Biochemistry (DCB) from Faculty of Sciences, University of Porto(FCUP), Porto, Portugal, where currently he is researcher. J.M.R. acknowledges the financial support of the strategic programmes UID/BIA/04050/2013 (POCI-01-0145-FEDER-007569) and PTDC/SAUNUT/30448/2017 (POCI-01-0145-FEDER-030448) and M.F.B. the financial support of the projects PTDC/QUI-QAN/30735/2017 and UID/QUI/50006/2019, funded by national funds through Fundação para a Ciência e Tecnologia [Foundation for Science and Technology](FCT-I.P.)/Ministério da Ciência, Tecnologia e Ensino Superior [Ministry of Science, Technology and Higher Education] (MCTES), and Fundo Europeu de Desenvolvimento Regional [European RegionalDevelopment Fund] (FEDER), under the scope of the COMPETE2020–Programa Operacional Competitividade e Internacionalização[COMPETE2020–Competitiveness and InternationalizationOperational Program, POCI]. S.B. acknowledges the financial support from MINECO through the“Juan de la Cierva” program.pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.doi10.1016/j.bios.2019.05.011pt_PT
dc.identifier.issn0956-5663
dc.identifier.urihttp://hdl.handle.net/10400.22/16004
dc.language.isoengpt_PT
dc.publisherElsevierpt_PT
dc.relationPTDC/SAUNUT/30448/2017pt_PT
dc.relation.publisherversionhttps://www.sciencedirect.com/science/article/pii/S0956566319303719?via%3Dihubpt_PT
dc.subjectSola l 7 allergen coding-sequencespt_PT
dc.subjectSandwich hybridizationpt_PT
dc.subjectAbRNA/DNApt_PT
dc.subjectScreen-printed electrodespt_PT
dc.subjectAmperometrypt_PT
dc.subjectGenomic DNA extractpt_PT
dc.titleDirect PCR-free electrochemical biosensing of plant-food derived nucleic acids in genomic DNA extracts. Application to the determination of the key allergen Sola l 7 in tomato seedspt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/5876/UID%2FBIA%2F04050%2F2013/PT
oaire.citation.endPage177pt_PT
oaire.citation.startPage171pt_PT
oaire.citation.titleBiosensors and Bioelectronicspt_PT
oaire.citation.volume137pt_PT
oaire.fundingStream5876
person.familyNamede Sá Barroso
person.givenNameMaria de Fátima
person.identifier.ciencia-idAF16-22E5-5EBD
person.identifier.orcid0000-0001-9011-5992
person.identifier.scopus-author-id25631648000
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.nameFundação para a Ciência e a Tecnologia
rcaap.rightsopenAccesspt_PT
rcaap.typearticlept_PT
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relation.isAuthorOfPublication.latestForDiscovery3d701c85-4580-4614-be6d-5a1abc8cf729
relation.isProjectOfPublication34715e6f-7ccc-439f-9322-558d880085d0
relation.isProjectOfPublication.latestForDiscovery34715e6f-7ccc-439f-9322-558d880085d0

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