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.

Design of a rapid, multiplex, one-pot miRNA assay optimized by label-free analysis / G. Zanchetta, T. Carzaniga, L. Vanjur, L. Casiraghi, G. Tagliabue, C. Morasso, T. Bellini, M. Buscaglia. - In: BIOSENSORS & BIOELECTRONICS. - ISSN 0956-5663. - 172(2021 Jan 15). [10.1016/j.bios.2020.112751]

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

G. Zanchetta
Primo
;
T. Carzaniga
Secondo
;
L. Vanjur;L. Casiraghi;G. Tagliabue;C. Morasso;T. Bellini
Penultimo
;
M. Buscaglia
Ultimo
2021-01-15

Abstract

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.
Label-free biosensor; Reflective phantom interface; Nucleic acids; Antibody; microRNA; DNA microarray; Rapid detection; Wash-free assay;
Settore FIS/07 - Fisica Applicata(Beni Culturali, Ambientali, Biol.e Medicin)
Piattaforma per l’identificazione di target di rilevanza farmacologica per il trattamento di patologie del sistema nervoso e oncologiche ad elevato bisogno di cura (NeOn)
ott-2020
Article (author)
File in questo prodotto:
File Dimensione Formato  
1-s2.0-S0956566320307399-main.pdf

accesso riservato

Descrizione: Articolo pubblicato
Tipologia: Publisher's version/PDF
Dimensione 3.47 MB
Formato Adobe PDF
3.47 MB Adobe PDF   Visualizza/Apri   Richiedi una copia
Zanchetta et al with SI.pdf

embargo fino al 15/01/2023

Descrizione: Articolo completo con supporting information
Tipologia: Post-print, accepted manuscript ecc. (versione accettata dall'editore)
Dimensione 1.95 MB
Formato Adobe PDF
1.95 MB Adobe PDF   Visualizza/Apri   Richiedi una copia
Pubblicazioni consigliate

Caricamento pubblicazioni consigliate

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/2434/781675
Citazioni
  • ???jsp.display-item.citation.pmc??? 3
  • Scopus 4
  • ???jsp.display-item.citation.isi??? 3
social impact