Autor según el artículo: J.L. Acero Sánchez; O.Y.F. Henry; H. Joda; B. Werne Solnestam; L. Kvastad; E. Johansson; P. Akan; J. Lundeberg; N. Lladach; D. Ramakrishnan; I. Riley; C.K. O'Sullivan
Departamento: Enginyeria Química
Autor/es de la URV: ACERO SÁNCHEZ, JOSEP LLUÍS; HENRY ., OLIVIER; H. Joda; B. Werne Solnestam; L. Kvastad; E. Johansson; P. Akan; J. Lundeberg; N. Lladach; D. Ramakrishnan; I. Riley; O'SULLIVAN ., CIARA
Palabras clave: Engineering controlled terms Electrochemical detection Engineering main heading
Resumen: Asymmetric multiplex ligation-dependent probe amplification (MLPA) was developed for the amplification of seven breast cancer related mRNA markers and the MLPA products were electrochemically detected via hybridization. Seven breast cancer genetic markers were amplified by means of the MLPA reaction, which allows for multiplex amplification of multiple targets with a single primer pair. Novel synthetic MLPA probes were designed to include a unique barcode sequence in each amplified gene. Capture probes complementary to each of the barcode sequences were immobilized on each electrode of a low-cost electrode microarray manufactured on standard printed circuit board (PCB) substrates. The functionalised electrodes were exposed to the single-stranded MLPA products and following hybridization, a horseradish peroxidase (HRP)-labelled DNA secondary probe complementary to the amplified strand completed the genocomplex, which was electrochemically detected following substrate addition. The electrode arrays fabricated using PCB technology exhibited an excellent electrochemical performance, equivalent to planar photolithographically-fabricated gold electrodes, but at a vastly reduced cost (>50 times lower per array). The optimised system was demonstrated to be highly specific with negligible cross-reactivity allowing the simultaneous detection of the seven mRNA markers, with limits of detections as low as 25 pM. This approach provides a novel strategy for the genetic profiling of tumour cells via integrated "amplification-to-detection".
Grupo de investigación: Group of Nanobiotechnology and Bioanalysis
Áreas temáticas: Enginyeria química Ingeniería química Chemical engineering
Acceso a la licencia de uso: https://creativecommons.org/licenses/by/3.0/es/
ISSN: 0956-5663
Identificador del autor: n/a; n/a; n/a; n/a; n/a; n/a; n/a; n/a; n/a; n/a; n/a; 0000-0002-0965-4655
Fecha de alta del registro: 2016-05-03
Página final: 232
Volumen de revista: 82
Versión del articulo depositado: info:eu-repo/semantics/acceptedVersion
Enlace a la fuente original: https://www.sciencedirect.com/science/article/pii/S0956566316302925?via%3Dihub
URL Documento de licencia: https://repositori.urv.cat/ca/proteccio-de-dades/
DOI del artículo: 10.1016/j.bios.2016.04.018
Entidad: Universitat Rovira i Virgili
Año de publicación de la revista: 2016
Página inicial: 224
Tipo de publicación: Article Artículo Article