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

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