CASCADE: Naked eye-detection of SARS-CoV-2 using Cas13a and gold nanoparticles.

Autor: López-Valls M; Fundación IMDEA Nanociencia, Madrid, Spain., Escalona-Noguero C; Fundación IMDEA Nanociencia, Madrid, Spain., Rodríguez-Díaz C; Fundación IMDEA Nanociencia, Madrid, Spain., Pardo D; Fundación IMDEA Nanociencia, Madrid, Spain; Synthelia Organics, Madrid, Spain., Castellanos M; Fundación IMDEA Nanociencia, Madrid, Spain., Milán-Rois P; Fundación IMDEA Nanociencia, Madrid, Spain., Martínez-Garay C; Fundación IMDEA Nanociencia, Madrid, Spain., Coloma R; Fundación IMDEA Nanociencia, Madrid, Spain., Abreu M; Servicio de Microbiología, Hospital Universitario Ramón y Cajal and Instituto Ramón y Cajal de Investigación Sanitaria (IRYCIS), 28034, Madrid, Spain., Cantón R; Servicio de Microbiología, Hospital Universitario Ramón y Cajal and Instituto Ramón y Cajal de Investigación Sanitaria (IRYCIS), 28034, Madrid, Spain; Red Española de Investigación en Patología Infecciosa (REIPI), Instituto de Salud Carlos III. Madrid, Spain., Galán JC; Servicio de Microbiología, Hospital Universitario Ramón y Cajal and Instituto Ramón y Cajal de Investigación Sanitaria (IRYCIS), 28034, Madrid, Spain; Centro de Investigación Biomédica en Red (CIBER) en Epidemiología y Salud Pública, Instituto de Salud Carlos III. Madrid, Spain., Miranda R; Fundación IMDEA Nanociencia, Madrid, Spain; Departamento de Física de la Materia Condensada, Universidad Autónoma de Madrid, Madrid, Spain., Somoza Á; Fundación IMDEA Nanociencia, Madrid, Spain; Nanobiotecnología (IMDEA-Nanociencia), Unidad Asociada al Centro Nacional de Biotecnología (CSIC), Madrid, Spain. Electronic address: alvaro.somoza@imdea.org., Sot B; Fundación IMDEA Nanociencia, Madrid, Spain; Nanobiotecnología (IMDEA-Nanociencia), Unidad Asociada al Centro Nacional de Biotecnología (CSIC), Madrid, Spain; Unidad de Innovación Biomédica, Centro de investigaciones Energéticas, Medioambientales y Tecnológicas (CIEMAT), Madrid, Spain. Electronic address: begona.sot@imdea.org.
Jazyk: angličtina
Zdroj: Analytica chimica acta [Anal Chim Acta] 2022 May 01; Vol. 1205, pp. 339749. Date of Electronic Publication: 2022 Mar 22.
DOI: 10.1016/j.aca.2022.339749
Abstrakt: The COVID-19 pandemic has brought to light the need for fast and sensitive detection methods to prevent the spread of pathogens. The scientific community is making a great effort to design new molecular detection methods suitable for fast point-of-care applications. In this regard, a variety of approaches have been developed or optimized, including isothermal amplification of viral nucleic acids, CRISPR-mediated target recognition, and read-out systems based on nanomaterials. Herein, we present CASCADE (CRISPR/CAS-based Colorimetric nucleic Acid DEtection), a sensing system for fast and specific naked-eye detection of SARS-CoV-2 RNA. In this approach, viral RNA is recognized by the LwaCas13a CRISPR protein, which activates its collateral RNase activity. Upon target recognition, Cas13a cleaves ssRNA oligonucleotides conjugated to gold nanoparticles (AuNPs), thus inducing their colloidal aggregation, which can be easily visualized. After an exhaustive optimization of functionalized AuNPs, CASCADE can detect picomolar concentrations of SARS-CoV-2 RNA. This sensitivity is further increased to low femtomolar (3 fM) and even attomolar (40 aM) ranges when CASCADE is coupled to RPA or NASBA isothermal nucleic acid amplification, respectively. We finally demonstrate that CASCADE succeeds in detecting SARS-CoV-2 in clinical samples from nasopharyngeal swabs. In conclusion, CASCADE is a fast and versatile RNA biosensor that can be coupled to different isothermal nucleic acid amplification methods for naked-eye diagnosis of infectious diseases.
Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(Copyright © 2022 The Authors. Published by Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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