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

doi:10.1016/j.bios.2020.112751

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

Research output: Contribution to journal › Article › peer-review

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.

Original language	English
Pages (from-to)	112751
Journal	Biosensors and Bioelectronics
Volume	172
DOIs	https://doi.org/10.1016/j.bios.2020.112751
Publication status	Published - Jan 15 2021

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

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.",

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AU - Zanchetta, G

AU - Carzaniga, T

AU - Vanjur, L

AU - Casiraghi, L

AU - Tagliabue, G

AU - Morasso, C

AU - Bellini, T

AU - Buscaglia, M

PY - 2021/1/15

Y1 - 2021/1/15

N2 - 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.

AB - 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.

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DO - 10.1016/j.bios.2020.112751

M3 - Article

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VL - 172

SP - 112751

JO - Biosensors and Bioelectronics

JF - Biosensors and Bioelectronics

ER -

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

Abstract

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