Exploring electrochemical impedance spectroscopy for the early diagnosis of Mycobacterium tuberculosis using CFP10:ESAT6 protein detection

Tuberculosis (TB) was, until SARS-CoV-2 pandemic, the leading cause of death by a single infectious agent contaminating over 10.6 million people with 1.6 million deaths in 2021 worldwide. Herein, we present a proof-of-principle strategy for detecting the recombinant protein CFP10:ESAT6 using an impe...

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Main Authors: Luisa Vogado Ribeiro, Juliana Cancino-Bernardi, Claudia do Amaral Razzino, Thales Rafael Machado, Marco A. M. Tuesta, Valtencir Zucolotto
Format: Article
Language:English
Published: Frontiers Media S.A. 2024-12-01
Series:Frontiers in Sensors
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Online Access:https://www.frontiersin.org/articles/10.3389/fsens.2024.1512936/full
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author Luisa Vogado Ribeiro
Juliana Cancino-Bernardi
Juliana Cancino-Bernardi
Claudia do Amaral Razzino
Thales Rafael Machado
Marco A. M. Tuesta
Valtencir Zucolotto
author_facet Luisa Vogado Ribeiro
Juliana Cancino-Bernardi
Juliana Cancino-Bernardi
Claudia do Amaral Razzino
Thales Rafael Machado
Marco A. M. Tuesta
Valtencir Zucolotto
author_sort Luisa Vogado Ribeiro
collection DOAJ
description Tuberculosis (TB) was, until SARS-CoV-2 pandemic, the leading cause of death by a single infectious agent contaminating over 10.6 million people with 1.6 million deaths in 2021 worldwide. Herein, we present a proof-of-principle strategy for detecting the recombinant protein CFP10:ESAT6 using an impedimetric immunosensor, which could aid in the diagnosis of tuberculosis. The immunosensor was developed using indium tin oxide electrodes modified by 3-aminopropyltrimethoxysilane monolayer to covalently immobilize anti-CFP10 antibodies. The protein interaction with the antibody recognition platform was directly monitored and measured by cyclic voltammetry and electrochemical impedance spectroscopy, respectively. After the analytical features optimization, a Langmuir isotherm response from 0.5 ng mL-1 to 50 ng mL-1 of pCFP10:ESAT6, limit of detection of 4.80 ng mL-1 and limit of quantification of 15.97 ng mL-1 were achieved, in a 4-hour assay time. Selectivity tests conducted in the presence of DENV NS1 and SARS-CoV-2 Spike proteins at a concentration of 20 ng mL-1, which is one-tenth of the concentration used to optimize pCFP10, indicate that the immunosensor is selective for pCFP10:ESAT6. Additionally, repeatability and reproducibility tests confirm that the immunosensor is suitable, accurate, and selective for detecting the CFP10:ESAT6 protein. The small sample volume required, and short testing time underscore the remarkable capabilities of this immunosensor and its potential for point-of-care screening and diagnostic aid applications.
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spelling doaj-art-457e224a620f47bcb0c8c912f7e17c2b2024-12-24T06:37:10ZengFrontiers Media S.A.Frontiers in Sensors2673-50672024-12-01510.3389/fsens.2024.15129361512936Exploring electrochemical impedance spectroscopy for the early diagnosis of Mycobacterium tuberculosis using CFP10:ESAT6 protein detectionLuisa Vogado Ribeiro0Juliana Cancino-Bernardi1Juliana Cancino-Bernardi2Claudia do Amaral Razzino3Thales Rafael Machado4Marco A. M. Tuesta5Valtencir Zucolotto6Nanomedicine and Nanotoxicology Group, Physics Institute of São Carlos, University of São Paulo, São Carlos, BrazilNanomedicine and Nanotoxicology Group, Physics Institute of São Carlos, University of São Paulo, São Carlos, BrazilChemistry Department, Laboratory in Bioanalytical of Nanosystems, Faculty of Philosophy, Science and Letters at Ribeirão Preto, University of São Paulo, Ribeirão Preto, BrazilNanomedicine and Nanotoxicology Group, Physics Institute of São Carlos, University of São Paulo, São Carlos, BrazilNanomedicine and Nanotoxicology Group, Physics Institute of São Carlos, University of São Paulo, São Carlos, BrazilNanomedicine and Nanotoxicology Group, Physics Institute of São Carlos, University of São Paulo, São Carlos, BrazilNanomedicine and Nanotoxicology Group, Physics Institute of São Carlos, University of São Paulo, São Carlos, BrazilTuberculosis (TB) was, until SARS-CoV-2 pandemic, the leading cause of death by a single infectious agent contaminating over 10.6 million people with 1.6 million deaths in 2021 worldwide. Herein, we present a proof-of-principle strategy for detecting the recombinant protein CFP10:ESAT6 using an impedimetric immunosensor, which could aid in the diagnosis of tuberculosis. The immunosensor was developed using indium tin oxide electrodes modified by 3-aminopropyltrimethoxysilane monolayer to covalently immobilize anti-CFP10 antibodies. The protein interaction with the antibody recognition platform was directly monitored and measured by cyclic voltammetry and electrochemical impedance spectroscopy, respectively. After the analytical features optimization, a Langmuir isotherm response from 0.5 ng mL-1 to 50 ng mL-1 of pCFP10:ESAT6, limit of detection of 4.80 ng mL-1 and limit of quantification of 15.97 ng mL-1 were achieved, in a 4-hour assay time. Selectivity tests conducted in the presence of DENV NS1 and SARS-CoV-2 Spike proteins at a concentration of 20 ng mL-1, which is one-tenth of the concentration used to optimize pCFP10, indicate that the immunosensor is selective for pCFP10:ESAT6. Additionally, repeatability and reproducibility tests confirm that the immunosensor is suitable, accurate, and selective for detecting the CFP10:ESAT6 protein. The small sample volume required, and short testing time underscore the remarkable capabilities of this immunosensor and its potential for point-of-care screening and diagnostic aid applications.https://www.frontiersin.org/articles/10.3389/fsens.2024.1512936/fullMycobacterium tuberculosisimmunosensorselectrochemical impedance spectroscopyimpedimetric biosensorsnanomedicine
spellingShingle Luisa Vogado Ribeiro
Juliana Cancino-Bernardi
Juliana Cancino-Bernardi
Claudia do Amaral Razzino
Thales Rafael Machado
Marco A. M. Tuesta
Valtencir Zucolotto
Exploring electrochemical impedance spectroscopy for the early diagnosis of Mycobacterium tuberculosis using CFP10:ESAT6 protein detection
Frontiers in Sensors
Mycobacterium tuberculosis
immunosensors
electrochemical impedance spectroscopy
impedimetric biosensors
nanomedicine
title Exploring electrochemical impedance spectroscopy for the early diagnosis of Mycobacterium tuberculosis using CFP10:ESAT6 protein detection
title_full Exploring electrochemical impedance spectroscopy for the early diagnosis of Mycobacterium tuberculosis using CFP10:ESAT6 protein detection
title_fullStr Exploring electrochemical impedance spectroscopy for the early diagnosis of Mycobacterium tuberculosis using CFP10:ESAT6 protein detection
title_full_unstemmed Exploring electrochemical impedance spectroscopy for the early diagnosis of Mycobacterium tuberculosis using CFP10:ESAT6 protein detection
title_short Exploring electrochemical impedance spectroscopy for the early diagnosis of Mycobacterium tuberculosis using CFP10:ESAT6 protein detection
title_sort exploring electrochemical impedance spectroscopy for the early diagnosis of mycobacterium tuberculosis using cfp10 esat6 protein detection
topic Mycobacterium tuberculosis
immunosensors
electrochemical impedance spectroscopy
impedimetric biosensors
nanomedicine
url https://www.frontiersin.org/articles/10.3389/fsens.2024.1512936/full
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