Articles producció científica> Enginyeria Química

Multiplex PCB-based electrochemical detection of cancer biomarkers using MLPA-barcode approach

  • Datos identificativos

    Identificador: PC:1527
    Autores:
    J.L. Acero SánchezO.Y.F. HenryH. JodaB. Werne SolnestamL. KvastadE. JohanssonP. AkanJ. LundebergN. LladachD. RamakrishnanI. RileyC.K. O'Sullivan
    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".
  • Otros:

    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
  • Palabras clave:

    Enginyeria química
    Sensors electroquímics
    Engineering controlled terms
    Electrochemical detection
    Engineering main heading
    Enginyeria química
    Ingeniería química
    Chemical engineering
    0956-5663
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