Author, as appears in the article.: 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
Department: Enginyeria Química
URV's Author/s: 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
Keywords: Engineering controlled terms Electrochemical detection Engineering main heading
Abstract: 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".
Research group: Group of Nanobiotechnology and Bioanalysis
Thematic Areas: Enginyeria química Ingeniería química Chemical engineering
licence for use: https://creativecommons.org/licenses/by/3.0/es/
ISSN: 0956-5663
Author identifier: 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
Record's date: 2016-05-03
Last page: 232
Journal volume: 82
Papper version: info:eu-repo/semantics/acceptedVersion
Link to the original source: https://www.sciencedirect.com/science/article/pii/S0956566316302925?via%3Dihub
Licence document URL: https://repositori.urv.cat/ca/proteccio-de-dades/
Article's DOI: 10.1016/j.bios.2016.04.018
Entity: Universitat Rovira i Virgili
Journal publication year: 2016
First page: 224
Publication Type: Article Artículo Article