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Doctoral Thesis
DOI
https://doi.org/10.11606/T.75.2023.tde-11122023-113549
Document
Author
Full name
Karla Patrícia Ribeiro Castro
Institute/School/College
Knowledge Area
Date of Defense
Published
São Carlos, 2023
Supervisor
Committee
Crespilho, Frank Nelson (President)
Cardoso, Daniel Rodrigues
Garcia, Marco Aurélio Suller
Kubota, Lauro Tatsuo
Moraes, Fernando Cruz de
Title in Portuguese
Desenvolvimento de sistema de quantificação de anticorpos contra o vírus SARS-CoV-2 utilizando imunoensaios de fluxo lateral
Keywords in Portuguese
monitoramento de anticorpo IgG
proteína RBD
SARS-CoV-2
teste de fluxo lateral
teste rápido
Abstract in Portuguese
Os imunoensaios de fluxo lateral (LFIA) modernizaram os testes em massa descentralizados e tiveram um impacto significativo no sistema de saúde. O teste sorológico é uma ferramenta importante para rastrear pacientes com fases virais passadas detectando anticorpos. Nessa tese de doutorado, apresenta-se o desenvolvimento de LFIA quantitativo, no qual os anticorpos anti-S-RBD presentes em amostras de soro de pacientes infectados previamente por SARS-CoV-2 formam um complexo imunológico com a proteína RBD marcada com nanopartículas de ouro. Para medidas quantitativas, foi desenvolvida uma metodologia baseada no uso de uma câmera digital que processa imagens das linhas de teste em câmara fechada. Os limites de detecção e quantificação foram 59, 5 ng mL-1 e 595,7 ng mL-1, respectivamente. O dispositivo proposto apresenta um custo de fabricação abaixo de US$ 1,0, desde a síntese dos bioconjugados, montagem das tiras LFIA e impressão 3D dos invólucros. A plataforma apresenta a capacidade de detectar anticorpos anti-S-RBD em amostras de soro humano podendo ser particularmente útil em estratégias de monitoramento de os níveis de IgG na população.
Title in English
Development quantification system for detection of antibody against the SARS-CoV-2 virus using lateral flow immunoassays
Keywords in English
lateral flow test
monitoring of IgG antibody
rapid test
RBD protein
SARS-CoV-2
Abstract in English
Lateral flow immunoassays (LFIA) have modernized decentralized mass testing and have had a significant impact on the healthcare system. Serological testing is an important tool to screen patients with past viral phases by detecting antibodies. In this doctoral thesis, the development of quantitative LFIA is presented, in which anti-S-RBD antibodies existing in serum samples from patients previously infected with SARS-CoV-2 form an immune complex with the RBD protein marked with nanoparticles of gold. For quantitative measurements, a methodology was developed based on the use of a digital camera that processes images of the test lines in a closed chamber. The limits of detection and quantification were 59.5 ng mL-1 and 595.7 ng mL-1, respectively. The proposed device has a manufacturing cost of less than US$ 1.0, from the synthesis of the bioconjugates, assembly of the LFIA strips and 3D printing of the envelopes. The platform could detect anti-S-RBD antibodies in human serum samples and can be particularly useful in strategies for monitoring IgG levels in the population.
 
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Release Date
2025-09-18
Publishing Date
2023-12-15
 
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