A genosensor for detecting single-point mutations in dendron chips after blocked recombinase polymerase amplification
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| dc.contributor.author | Martorell, Sara
|
es_ES |
| dc.contributor.author | Maquieira Catala, Angel
|
es_ES |
| dc.contributor.author | Tortajada-Genaro, Luis Antonio
|
es_ES |
| dc.date.accessioned | 2022-11-22T19:03:06Z | |
| dc.date.available | 2022-11-22T19:03:06Z | |
| dc.date.issued | 2022-05-17 | es_ES |
| dc.identifier.issn | 0003-2654 | es_ES |
| dc.identifier.uri | http://hdl.handle.net/10251/190065 | |
| dc.description.abstract | [EN] A biosensing system was developed to accurately detect a single nucleotide change in the target organism genome, integrating a selective isothermal amplification and a sensitive dendron-mediated DNA hybridization assay in the array format. The novelty arises from the coupling reactions of the dendron and its use as a crosslinker. The allele-specific probes were oligonucleotide-dendron conjugates prepared by fast and clean click-chemistry (thiol-yne reaction) and coupled onto the photo-activated surface of polycarbonate substrates (carbodiimide reaction). The output was forest-array chips with multipoint-site crosslinkers and compatible with current microarray fabrication technologies. The products of blocked recombinase polymerase amplification (blocked RPA), formed at 37 degrees C, were hybridized with attached probes for specific nucleotide genotyping. The developed approach exhibited sensitive recognition of DNA variants compared to chips based on linear crosslinkers (10-100 fold), showing excellent analytical performances for planar chip and fluidic formats. The methodology was successfully applied to detect the H1047R mutation in the PIK3CA gene (c.3140A > G) from clinical samples of human cancer tissues, the results being consistent with sequencing techniques. The colorimetric biosensing method was reliable, versatile, low cost, sensitive (detection limit genomic DNA: 0.02 ng mu L-1), and specific (accuracy >95%). | es_ES |
| dc.description.sponsorship | The MINECO Project PID2019-110713RB-I00 and Generalitat Valenciana PROMETEO/2020/094 are acknowledged. | es_ES |
| dc.language | Inglés | es_ES |
| dc.publisher | The Royal Society of Chemistry | es_ES |
| dc.relation.ispartof | The Analyst | es_ES |
| dc.rights | Reserva de todos los derechos | es_ES |
| dc.subject.classification | QUIMICA ANALITICA | es_ES |
| dc.title | A genosensor for detecting single-point mutations in dendron chips after blocked recombinase polymerase amplification | es_ES |
| dc.type | Artículo | es_ES |
| dc.identifier.doi | 10.1039/d2an00160h | es_ES |
| dc.relation.projectID | info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/PID2019-110713RB-I00/ES/HACIA EL BIOSENSADO HOLOGRAFICO AVANZADO/ | es_ES |
| dc.relation.projectID | info:eu-repo/grantAgreement/GENERALITAT VALENCIANA//PROMETEO%2F2020%2F094//HOLOGRAFÍA. UNA VIA PARA AFRONTAR NUEVOS RETOS EN BIOSENSADO/ | es_ES |
| dc.rights.accessRights | Abierto | es_ES |
| dc.contributor.affiliation | Universitat Politècnica de València. Escuela Técnica Superior de Ingeniería Agronómica y del Medio Natural - Escola Tècnica Superior d'Enginyeria Agronòmica i del Medi Natural | es_ES |
| dc.description.bibliographicCitation | Martorell, S.; Maquieira Catala, A.; Tortajada-Genaro, LA. (2022). A genosensor for detecting single-point mutations in dendron chips after blocked recombinase polymerase amplification. The Analyst. 147(10):2180-2188. https://doi.org/10.1039/d2an00160h | es_ES |
| dc.description.accrualMethod | S | es_ES |
| dc.relation.publisherversion | https://doi.org/10.1039/d2an00160h | es_ES |
| dc.description.upvformatpinicio | 2180 | es_ES |
| dc.description.upvformatpfin | 2188 | es_ES |
| dc.type.version | info:eu-repo/semantics/publishedVersion | es_ES |
| dc.description.volume | 147 | es_ES |
| dc.description.issue | 10 | es_ES |
| dc.identifier.pmcid | 35441629 | es_ES |
| dc.relation.pasarela | S\463971 | es_ES |
| dc.contributor.funder | GENERALITAT VALENCIANA | es_ES |
| dc.contributor.funder | AGENCIA ESTATAL DE INVESTIGACION | es_ES |
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| dc.subject.ods | 03.- Garantizar una vida saludable y promover el bienestar para todos y todas en todas las edades | es_ES |
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