Design of a rapid, multiplex, one-pot miRNA assay optimized by label-free analysis.

Biological Assay Biomarkers Biosensing Techniques DNA Probes MicroRNAs
Antibody DNA microarray Label-free biosensor Nucleic acids Rapid detection Reflective phantom interface Wash-free assay microRNA

Journal

Biosensors & bioelectronics
ISSN: 1873-4235
Titre abrégé: Biosens Bioelectron
Pays: England
ID NLM: 9001289

Informations de publication

Date de publication:
15 Jan 2021
Historique:
received: 22 07 2020
revised: 06 10 2020
accepted: 20 10 2020
pubmed: 3 11 2020
medline: 15 5 2021
entrez: 2 11 2020
Statut: ppublish

Résumé

MicroRNAs are widely studied as circulating biomarkers for early stage diagnosis of several diseases. Detection and quantification of miRNAs is currently performed through complex and time consuming procedures. Herein we demonstrate a rapid, multiplex, one-pot detection method based on two-step amplification of the signal measured by Reflective Phantom Interface (RPI) label-free optical biosensor. We achieved sub-pM quantification of different miRNAs in about 1.5 h, through specific capture with surface DNA probes combined to a 35-fold mass amplification by an antibody targeting DNA-RNA hybrids and polyclonal secondary antibody, all performed without washing steps. The assay is the result of a modelling and optimization of the multi-step process that has been made possible by the RPI characterization of each individual interaction involved.

Identifiants

DOI: 10.1016/j.bios.2020.112751 PMID: 33137609
pubmed: 33137609
pii: S0956-5663(20)30739-9
doi: 10.1016/j.bios.2020.112751
pii:
doi:

Substances chimiques

Biomarkers 0
DNA Probes 0
MicroRNAs 0

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

112751

Informations de copyright

Copyright © 2020 Elsevier B.V. All rights reserved.

Auteurs

G Zanchetta (G)

Dipartimento di Biotecnologie Mediche e Medicina Traslazionale, Università degli Studi di Milano, Segrate, Italy.

T Carzaniga (T)

Dipartimento di Biotecnologie Mediche e Medicina Traslazionale, Università degli Studi di Milano, Segrate, Italy.

L Vanjur (L)

Dipartimento di Biotecnologie Mediche e Medicina Traslazionale, Università degli Studi di Milano, Segrate, Italy.

L Casiraghi (L)

Dipartimento di Biotecnologie Mediche e Medicina Traslazionale, Università degli Studi di Milano, Segrate, Italy.

G Tagliabue (G)

Proxentia S.r.l., viale Ortles 22/4, Milano, Italy.

C Morasso (C)

Nanomedicine and Molecular Imaging Laboratory, Istituti Clinici Scientifci Maugeri IRCCS, Pavia, Italy.

T Bellini (T)

Dipartimento di Biotecnologie Mediche e Medicina Traslazionale, Università degli Studi di Milano, Segrate, Italy.

M Buscaglia (M)

Dipartimento di Biotecnologie Mediche e Medicina Traslazionale, Università degli Studi di Milano, Segrate, Italy. Electronic address: marco.buscaglia@unimi.it.

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Classifications MeSH

questionsmedicales.fr › Techniques d'investigation › Dosage biologique › Design of a rapid, multiplex, one-pot miRNA...
questionsmedicales.fr › Facteurs biologiques › Marqueurs biologiques › Design of a rapid, multiplex, one-pot miRNA...
questionsmedicales.fr › Techniques d'investigation › Techniques de sonde moléculaire › Techniques de biocapteur › Design of a rapid, multiplex, one-pot miRNA...
questionsmedicales.fr › Actions chimiques et utilisations › Utilisations spécialisées de produits chimiques › Produits chimiques de laboratoire › Sondes moléculaires › Sondes d'acide nucléique › Sondes d'ADN › Design of a rapid, multiplex, one-pot miRNA...
questionsmedicales.fr › Acides nucléiques, nucléotides et nucléosides › Acides nucléiques › ARN › ARN non traduit › Petit ARN non traduit › microARN › Design of a rapid, multiplex, one-pot miRNA...