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Master's Dissertation
DOI
https://doi.org/10.11606/D.76.2021.tde-10112021-094941
Document
Author
Full name
Gustavo Freitas do Nascimento
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Carlos, 2021
Supervisor
Committee
Oliveira Junior, Osvaldo Novais de (President)
Júnior, Antonio Riul
Yamanaka, Hideko
Title in Portuguese
Desenvolvimento de dispositivos para o monitoramento da bactéria Fusobacterium nucleatum para o diagnóstico precoce de câncer colorretal
Keywords in Portuguese
Câncer colorretal
Eletrodos impressos
Fusobacterium nucleatum
Genossensor
Micropartículas magnéticas
Abstract in Portuguese
Monitorar a presença de microrganismos em fluidos corporais é essencial para o diagnóstico de diversas doenças, como é o caso da Fusobacterium nucleatum, associada ao câncer colorretal. Para que esse monitoramento seja de baixo custo e implementável em sistemas públicos de saúde, novas metodologias precisam ser desenvolvidas. Neste trabalho, foram produzidos genossensores eletroquímicos com eletrodos de carbono impressos com micropartículas magnéticas para pré-concentrar a sequência de DNA da Fusobacterium nucleatum. Em condições otimizadas com voltametria de onda quadrada, o genossensor apresentou resposta sigmoidal representada pela isoterma de adsorção de Langmuir-Freundlich para a sequência complementar (target) nas concentrações de 1,38 a 172,5 μg mL⁻, com limite de detecção e de quantificação de 2,03 μg mL⁻1 e 6,78 μg mL⁻1, respectivamente. Esse genossensor também foi capaz de detectar a sequência target em extrato celular de linhagens de células (SW620 e HT-29) de pacientes com câncer colorretal, com taxa de recuperação de 85 a 108%. Conclui-se que a metodologia proposta é uma alternativa para o diagnóstico e prognóstico de baixo-custo e não invasivo de câncer colorretal.
Title in English
Low-cost disposable screen-printed carbon based eletrochemical device for early diagnosis of colorectal cancer
Keywords in English
Biosensor
Colorectal cancer
Dynabeads
Fusobacterium nucleatum
Genosensor
Screen-printed electrodes
Abstract in English
Monitoring the presence of microorganisms in body fluids is essential for the early diagnosis of various disorders, as in the case of Fusobacterium nucleatum that is associated with colorectal cancer. If this monitoring is to be implemented in large scale in the public health system, low-cost methodologies need to be developed. In this work, we produced eletrochemical genosensors made with screen-printed carbon electrodes which are employed with magnetic dynabeads to pre-concentrate and detect Fusobacterium nucleatum. Under optimized conditions, the genosensor exhibited a sigmoidal response in square wave voltammetry for the complementary sequence (target) of Fusobacterium nucleatum from 1.38 to 172.5 μg mL⁻1 , with a detection limit of 2.03 μg mL⁻1 and limit of quantification of 6.78 μg mL⁻1 . This genosensor was also capable of detecting the target in cell lysates (SW620 and HT-29), with recovery from 85 to 108%. The proposed methodology may represent an efficient, alternative method for low-cost and non-invasive diagnostic and prognostic of colorectal cancer.
 
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Publishing Date
2021-11-11
 
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