Pushing the Limits of Lateral Flow Immunoassay by Digital SERS for the Ultralow Detection of SARS‐CoV‐2 Virus

Lateral flow immunoassays (LFIAs) are easy‐to‐use antigen tests that provide different signal readouts, with colorimetric readouts being the most commonly used. However, these analytical devices have relatively low sensitivity and produce semiquantitative results, limiting their diagnostic applicati...

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Main Authors: Lara González‐Cabaleiro, Carlos Fernández‐Lodeiro, Lorena Vázquez‐Iglesias, Pablo Soriano‐Maldonado, Mark J. van Raaij, Gustavo Bodelón, Jorge Pérez‐Juste, Isabel Pastoriza‐Santos
Format: Article
Language:English
Published: Wiley-VCH 2024-11-01
Series:Small Science
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Online Access:https://doi.org/10.1002/smsc.202400259
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author Lara González‐Cabaleiro
Carlos Fernández‐Lodeiro
Lorena Vázquez‐Iglesias
Pablo Soriano‐Maldonado
Mark J. van Raaij
Gustavo Bodelón
Jorge Pérez‐Juste
Isabel Pastoriza‐Santos
author_facet Lara González‐Cabaleiro
Carlos Fernández‐Lodeiro
Lorena Vázquez‐Iglesias
Pablo Soriano‐Maldonado
Mark J. van Raaij
Gustavo Bodelón
Jorge Pérez‐Juste
Isabel Pastoriza‐Santos
author_sort Lara González‐Cabaleiro
collection DOAJ
description Lateral flow immunoassays (LFIAs) are easy‐to‐use antigen tests that provide different signal readouts, with colorimetric readouts being the most commonly used. However, these analytical devices have relatively low sensitivity and produce semiquantitative results, limiting their diagnostic applications. Herein, we address these challenges by implementing a digital surface‐enhanced Raman spectroscopy (SERS)‐based LFIA for the accurate and ultrasensitive quantitative detection of SARS‐CoV‐2 nucleocapsid (N) protein. Compared with average SERS intensity measurements, the digital approach allowed to overcome fluctuations in Raman scattering signals, thereby increasing the analytical sensitivity of the assay. Our method exhibited a quantification range of the viral protein in nasal swabs from 0.001 to 10 pg mL−1, and a limit of detection down to 1.9 aM (0.9 fg mL−1), improving colorimetric LFIAs and conventional‐SERS‐based LFIAs by several orders of magnitude. Importantly, this approach shows an analytical sensitivity of 0.03 TCID50 mL−1, which is greater than that reported by other immunoassays. In conclusion, we successfully demonstrate the robust detection and quantification of SARS‐CoV‐2N protein in nasal swabs at ultralow concentrations. The improvement in the sensitivity of LFIA by digital SERS may pave the way to translate this technology into the diagnostic arena for the ultrasensitive detection of microbes and disease biomarkers.
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spelling doaj-art-489cfb48c91a4632957cf307abe3731c2024-11-11T15:33:38ZengWiley-VCHSmall Science2688-40462024-11-01411n/an/a10.1002/smsc.202400259Pushing the Limits of Lateral Flow Immunoassay by Digital SERS for the Ultralow Detection of SARS‐CoV‐2 VirusLara González‐Cabaleiro0Carlos Fernández‐Lodeiro1Lorena Vázquez‐Iglesias2Pablo Soriano‐Maldonado3Mark J. van Raaij4Gustavo Bodelón5Jorge Pérez‐Juste6Isabel Pastoriza‐Santos7CINBIO Universidade de Vigo, Campus Universitario As Lagoas Marcosende Vigo 36310 SpainCINBIO Universidade de Vigo, Campus Universitario As Lagoas Marcosende Vigo 36310 SpainCINBIO Universidade de Vigo, Campus Universitario As Lagoas Marcosende Vigo 36310 SpainCentro Nacional de Biotecnología (CNB‐CSIC) calle Darwin 3 Madrid 28049 SpainCentro Nacional de Biotecnología (CNB‐CSIC) calle Darwin 3 Madrid 28049 SpainCINBIO Universidade de Vigo, Campus Universitario As Lagoas Marcosende Vigo 36310 SpainCINBIO Universidade de Vigo, Campus Universitario As Lagoas Marcosende Vigo 36310 SpainCINBIO Universidade de Vigo, Campus Universitario As Lagoas Marcosende Vigo 36310 SpainLateral flow immunoassays (LFIAs) are easy‐to‐use antigen tests that provide different signal readouts, with colorimetric readouts being the most commonly used. However, these analytical devices have relatively low sensitivity and produce semiquantitative results, limiting their diagnostic applications. Herein, we address these challenges by implementing a digital surface‐enhanced Raman spectroscopy (SERS)‐based LFIA for the accurate and ultrasensitive quantitative detection of SARS‐CoV‐2 nucleocapsid (N) protein. Compared with average SERS intensity measurements, the digital approach allowed to overcome fluctuations in Raman scattering signals, thereby increasing the analytical sensitivity of the assay. Our method exhibited a quantification range of the viral protein in nasal swabs from 0.001 to 10 pg mL−1, and a limit of detection down to 1.9 aM (0.9 fg mL−1), improving colorimetric LFIAs and conventional‐SERS‐based LFIAs by several orders of magnitude. Importantly, this approach shows an analytical sensitivity of 0.03 TCID50 mL−1, which is greater than that reported by other immunoassays. In conclusion, we successfully demonstrate the robust detection and quantification of SARS‐CoV‐2N protein in nasal swabs at ultralow concentrations. The improvement in the sensitivity of LFIA by digital SERS may pave the way to translate this technology into the diagnostic arena for the ultrasensitive detection of microbes and disease biomarkers.https://doi.org/10.1002/smsc.202400259digital SERSNanoBiosensorSARS‐CoV‐2SERS‐based LFIA
spellingShingle Lara González‐Cabaleiro
Carlos Fernández‐Lodeiro
Lorena Vázquez‐Iglesias
Pablo Soriano‐Maldonado
Mark J. van Raaij
Gustavo Bodelón
Jorge Pérez‐Juste
Isabel Pastoriza‐Santos
Pushing the Limits of Lateral Flow Immunoassay by Digital SERS for the Ultralow Detection of SARS‐CoV‐2 Virus
Small Science
digital SERS
NanoBiosensor
SARS‐CoV‐2
SERS‐based LFIA
title Pushing the Limits of Lateral Flow Immunoassay by Digital SERS for the Ultralow Detection of SARS‐CoV‐2 Virus
title_full Pushing the Limits of Lateral Flow Immunoassay by Digital SERS for the Ultralow Detection of SARS‐CoV‐2 Virus
title_fullStr Pushing the Limits of Lateral Flow Immunoassay by Digital SERS for the Ultralow Detection of SARS‐CoV‐2 Virus
title_full_unstemmed Pushing the Limits of Lateral Flow Immunoassay by Digital SERS for the Ultralow Detection of SARS‐CoV‐2 Virus
title_short Pushing the Limits of Lateral Flow Immunoassay by Digital SERS for the Ultralow Detection of SARS‐CoV‐2 Virus
title_sort pushing the limits of lateral flow immunoassay by digital sers for the ultralow detection of sars cov 2 virus
topic digital SERS
NanoBiosensor
SARS‐CoV‐2
SERS‐based LFIA
url https://doi.org/10.1002/smsc.202400259
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