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Campo DC | Valor | Lengua/Idioma |
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dc.contributor.author | Loaiza, Óscar A. | - |
dc.contributor.author | Campuzano, Susana | - |
dc.contributor.author | Pedrero, María | - |
dc.contributor.author | Pividori, M. Isabel | - |
dc.contributor.author | García, Pedro | - |
dc.contributor.author | Pingarrón, José Manuel | - |
dc.date.accessioned | 2012-09-25T11:12:31Z | - |
dc.date.available | 2012-09-25T11:12:31Z | - |
dc.date.issued | 2008-11-01 | - |
dc.identifier.citation | Analytical Chemistry 80(21):8239-8245(2008) | es_ES |
dc.identifier.issn | 0003-2700 | - |
dc.identifier.uri | http://hdl.handle.net/10261/56682 | - |
dc.description | 7 páginas, 8 figuras, 1 tabla -- PAGS nros. 8239-8245 | es_ES |
dc.description.abstract | Disposable magnetic DNA sensors using an enzyme-amplified strategy for the specific detection of a gene related to the Enterobacteriaceae bacterial family, based on the coupling of streptavidin-peroxidase to biotinylated lacZ gene target sequences, has been developed. A biotinylated 25-mer capture probe was attached to streptavidin-modified magnetic beads and hybridization with the biotinylated target was allowed to proceed. Then, a streptavidin-peroxidase polymer was attached to the biotinylated target, and the resulting modified magnetic beads were captured by a magnetic field on the surface of tetrathiafulvalene (TTF) modified gold screen-printed electrodes (Au/SPEs). The amperometric response obtained at −0.15 V after the addition of hydrogen peroxide was used to detect the hybridization process. In order to improve the sensitivity of the determination and reduce the assay time, different variables of the assay protocol were optimized. A low detection limit (5.7 fmol) with good stability (RSD = 7.1%, n = 10) was obtained. The DNA nonspecific adsorption at the magnetic beads was negligible, the obtained results thus demonstrating the possibility to detect the hybridization event with great specificity and sensitivity. The developed method was used for the analysis of Escherichia coli DNA fragments (326 bases) in polymerase chain reaction (PCR) amplicons extracted from a cell culture. As low as 2.5 aM asymmetric PCR product could be detected with the developed methodology | es_ES |
dc.description.sponsorship | The financial support of Santander/Complutense Research Project PR 27/05-13953 and of the Spanish Ministerio de Educación y Ciencia Research Project CTQ2006-02743BQU is gratefully acknowledged. Oscar A. Loaiza acknowledges a pre-Ph.D. fellowship of the Universidad Complutense de Madrid. Susana Campuzano acknowledges a “Juan de la Cierva” contract to the Spanish Ministerio de Educación y Ciencia | es_ES |
dc.language.iso | eng | es_ES |
dc.publisher | American Chemical Society | es_ES |
dc.rights | closedAccess | es_ES |
dc.title | Disposable magnetic DNA sensors for the determination at the attomolar level of a specific Enterobacteriaceae family gene | es_ES |
dc.type | artículo | es_ES |
dc.identifier.doi | 10.1021/ac801319b | - |
dc.description.peerreviewed | Peer reviewed | es_ES |
dc.relation.publisherversion | http://dx.doi.org/10.1021/ac801319b | es_ES |
dc.identifier.e-issn | 1520-6882 | - |
dc.type.coar | http://purl.org/coar/resource_type/c_6501 | es_ES |
item.openairecristype | http://purl.org/coar/resource_type/c_18cf | - |
item.fulltext | No Fulltext | - |
item.cerifentitytype | Publications | - |
item.openairetype | artículo | - |
item.languageiso639-1 | en | - |
item.grantfulltext | none | - |
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