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Parallel solid-phase isothermal amplification and detection of multiple DNA targets in microliter-sized wells of a digital versatile disc

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Parallel solid-phase isothermal amplification and detection of multiple DNA targets in microliter-sized wells of a digital versatile disc

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dc.contributor.author Santiago Felipe, Sara es_ES
dc.contributor.author Tortajada-Genaro, Luis Antonio es_ES
dc.contributor.author Puchades, Rosa es_ES
dc.contributor.author Maquieira Catala, Ángel es_ES
dc.date.accessioned 2016-09-12T06:47:33Z
dc.date.available 2016-09-12T06:47:33Z
dc.date.issued 2016-03
dc.identifier.issn 0026-3672
dc.identifier.uri http://hdl.handle.net/10251/69238
dc.description.abstract An integrated method for the parallelized detection of multiple DNA target sequences is presented by using microstructures in a digital versatile disc (DVD). Samples and reagents were managed by using both the capillary and centrifugal forces induced by disc rotation. Recombinase polymerase amplification (RPA), in a bridge solid phase format, took place in separate wells, which thereby modified their optical properties. Then the DVD drive reader recorded the modifications of the transmitted laser beam. The strategy allowed tens of genetic determinations to be made simultaneously within < 2 h, with small sample volumes (3 mu L), low manipulation and at low cost. The method was applied to high-throughput screening of relevant safety threats (allergens, GMOs and pathogenic bacteria) in food samples. Satisfactory results were obtained in terms of sensitivity (48.7 fg of DNA) and reproducibility (below 18 %). This scheme warrants cost-effective multiplex amplification and detection and is perceived to represent a viable tool for screening of nucleic acid targets. es_ES
dc.description.sponsorship This research has been funded through projects FEDER PrometeoII/2014/040 (GVA), and CTQ/2013/45875-R (MINECO). The Spanish Ministry of Education and Science provided S.S.F. with a grant for PhD studies. en_EN
dc.language Inglés es_ES
dc.publisher Springer Verlag (Germany) es_ES
dc.relation.ispartof Microchimica Acta es_ES
dc.rights Reserva de todos los derechos es_ES
dc.subject Multiplex PCR es_ES
dc.subject DNA es_ES
dc.subject Recombinase polymerase amplification (RPA) es_ES
dc.subject Compact disc es_ES
dc.subject High-throughput screening es_ES
dc.subject Food safety es_ES
dc.subject.classification QUIMICA ANALITICA es_ES
dc.title Parallel solid-phase isothermal amplification and detection of multiple DNA targets in microliter-sized wells of a digital versatile disc es_ES
dc.type Artículo es_ES
dc.identifier.doi 10.1007/s00604-016-1745-3
dc.relation.projectID info:eu-repo/grantAgreement/GVA//PROMETEOII%2F2014%2F040/ES/Estudio de estrategias fisico-químicas para el desarrollo de biosensores interferométricos en soportes interactivos de aplicación en clínica/ es_ES
dc.relation.projectID info:eu-repo/grantAgreement/MINECO//CTQ2013-45875-R/ES/BIOSENSADO EN SOPORTES INTERACTIVOS CON PROPIEDADES INTERFEROMETRICAS PARA APLICACIONES CLINICAS. DEMOSTRACION EN FARMACOGENETICA Y ALERGIAS MEDICAMENTOSAS/ es_ES
dc.rights.accessRights Abierto es_ES
dc.contributor.affiliation Universitat Politècnica de València. Departamento de Química - Departament de Química es_ES
dc.description.bibliographicCitation Santiago Felipe, S.; Tortajada-Genaro, LA.; Puchades, R.; Maquieira Catala, Á. (2016). Parallel solid-phase isothermal amplification and detection of multiple DNA targets in microliter-sized wells of a digital versatile disc. Microchimica Acta. 183(3):1195-1202. https://doi.org/10.1007/s00604-016-1745-3 es_ES
dc.description.accrualMethod S es_ES
dc.relation.publisherversion https://dx.doi.org/10.1007/s00604-016-1745-3 es_ES
dc.description.upvformatpinicio 1195 es_ES
dc.description.upvformatpfin 1202 es_ES
dc.type.version info:eu-repo/semantics/publishedVersion es_ES
dc.description.volume 183 es_ES
dc.description.issue 3 es_ES
dc.relation.senia 306763 es_ES
dc.identifier.eissn 1436-5073
dc.contributor.funder Generalitat Valenciana es_ES
dc.contributor.funder Ministerio de Economía y Competitividad es_ES
dc.contributor.funder Ministerio de Educación y Ciencia es_ES
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