LAMP-CRISPR/Cas12a-based impedimetric biosensor powered by Fe<sub>3</sub>O<sub>4</sub>@Au-(S-polyA-S)-Au for detection of SARS-CoV-2.

Behnam Rad, Mohammad; Hakimian, Fatemeh; Mohebbi, Seyed Reza; Yadegar, Abbas; Ghourchian, Hedayatollah

LAMP-CRISPR/Cas12a-based impedimetric biosensor powered by Fe₃O₄@Au-(S-polyA-S)-Au for detection of SARS-CoV-2.

Behnam Rad, Mohammad; Hakimian, Fatemeh; Mohebbi, Seyed Reza; Yadegar, Abbas; Ghourchian, Hedayatollah.

Afiliação

Behnam Rad M; Laboratory of Bioanalysis, Institute of Biochemistry & Biophysics, University of Tehran, Tehran, Iran.
Hakimian F; Laboratory of Bioanalysis, Institute of Biochemistry & Biophysics, University of Tehran, Tehran, Iran.
Mohebbi SR; Gastroenterology and Liver Diseases Research Center, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Yadegar A; Foodborne and Waterborne Diseases Research Center, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Ghourchian H; Laboratory of Bioanalysis, Institute of Biochemistry & Biophysics, University of Tehran, Tehran, Iran. ghourchian@ut.ac.ir.

Mikrochim Acta ; 191(11): 644, 2024 Oct 03.

Article em En | MEDLINE | ID: mdl-39361061

ABSTRACT

ABSTRACT

A low-cost, lab-made polytetrafluoroethylene micro-cell, equipped with three electrodes, wasd eveloped for the impedimetric detection of SARS-CoV-2. The gold working electrode was modified with a double-ended thiolated poly-adenine probe, which was conjugated with magnetic Fe3O4@Au nanoparticles (Fe3O4@Au-(S-polyA-S)-Au). After the loop-mediated isothermal amplification (LAMP) of viral RNA, the single-guide RNA (sgRNA), specifically bound to the SARS-CoV-2 target sequence, activates Cas12a. Cas12a then cleaved the immobilized probe. As a result, the magnetic Fe3O4@Au nanoparticles were released and adsorbed onto the gold electrode surface, using an external magnet. This process increased the physical surface area of the gold electrode, facilitating redox ion ([FeIII/II(CN)6]3-/4-) electron transfer. The decrease in the charge transfer resistance was utilized for SARS-CoV-2 detection. Our LAMP-CRISPR/Cas12a-based impedimetric biosensor, powered by Fe3O4@Au-(S-polyA-S)-Au, demonstrated impressive capabilities, including a remarkable detection limit of 0.8 aM (0.48 copies/µL) and a linear range of 0.01 to 36.06 fM.

Assuntos

Técnicas Biossensoriais; Sistemas CRISPR-Cas; Ouro; Técnicas de Amplificação de Ácido Nucleico; RNA Viral; SARS-CoV-2; SARS-CoV-2/genética; SARS-CoV-2/isolamento & purificação; Técnicas Biossensoriais/métodos; Ouro/química; Técnicas de Amplificação de Ácido Nucleico/métodos; Humanos; RNA Viral/análise; COVID-19/diagnóstico; COVID-19/virologia; Limite de Detecção; Eletrodos; Poli A/química; Proteínas Associadas a CRISPR; Nanopartículas de Magnetita/química; Endodesoxirribonucleases/química; Nanopartículas Metálicas/química; Proteínas de Bactérias; Técnicas de Diagnóstico Molecular

Palavras-chave

CRISPR/Cas; EIS ; Electrochemical biosensing; Modified gold electrode; Point-of-care; SARS-CoV-2

Texto completo

Adicionar na Minha BVS

Imprimir

XML

PubMed Links

Buscar no Google

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: RNA Viral / Técnicas Biossensoriais / Técnicas de Amplificação de Ácido Nucleico / Sistemas CRISPR-Cas / SARS-CoV-2 / Ouro Limite: Humans Idioma: En Revista: Mikrochim Acta Ano de publicação: 2024 Tipo de documento: Article País de afiliação: Irã País de publicação: Áustria

Texto completo

Adicionar na Minha BVS

Imprimir

XML

PubMed Links

Buscar no Google