A carregar...
Publicação
Análise e otimização da tecnologia de transdução ótica por ressonância de plasmão de superfície na determinação da concentração de Proteína C Reativa (PCR)
| Nome: | Descrição: | Tamanho: | Formato: | |
|---|---|---|---|---|
| 2.1 MB | Adobe PDF |
Orientador(es)
Resumo(s)
A tecnologia de point-of-care surgiu de forma a permitir o diagnóstico rápido e eficaz, tornando possível uma intervenção e tratamento atempados por parte do profissional de saúde. A Biosurfit S.A. é uma empresa portuguesa que desenvolve e produz dispositivos médicos que permitem a realização de diversos testes quantitativos perto do paciente. Além da máquina onde se processa a leitura de dados, o spinit®, existe um conjunto de discos multi-parmétricos descartáveis que, mediante a utilização de uma única gota de sangue e sem necessidade de reagentes adicionais por parte do utilizador, fornecem resultados quantitativos num período inferior a 15 minutos. Um dos testes desenvolvidos foi a análise da proteína C reativa, cujo disco possui várias zonas de deteção que incluem a camada de reconhecimento biológico, que contém fragmentos de anticorpos imobilizados numa superfície de ouro. Quando o plasma do paciente passa por estas zonas, o marcador-alvo é ligado aos fragmentos de anticorpos imobilizados. Este fenómeno induz uma alteração no índice de refração que, posteriormente, é amplificado pela ligação de nanopartículas de ouro, também estas específicas do analito. A tecnologia do leitor tem por base o fenómeno de ressonância de plasmão de superfície. Neste contexto, esta dissertação teve como objetivo inicial a redução do tempo de teste sem alteração da gama de deteção da proteína C reativa. Ao longo do estudo verificou-se que com a dimensão das nanopartículas de ouro existentes no sistema tornava-se inexequível o cumprimento do objetivo, pois a gama era diminuída. Tal foi possível através da utilização de nanopartículas de ouro de 20 nm que permitiram, para além da diminuição do tempo de teste (objetivo principal), aumentar a gama de deteção. Os resultados obtidos permitiram validar analiticamente a melhoria do produto e submeter uma certificação europeia do mesmo, uma vez que a deteção da proteína C reativa foi melhorada de 5 a 100 mg/L para 4 a 180 mg/L e o tempo de ensaio encurtado de 15 para 6 minutos.
Point-of-care technology emerged in order to allow a quick and efficient diagnosis, making possible a timely intervention and treatment by healthcare professionals. Biosurfit SA. is a company that developed and produced a medical equipment that allows to perform several quantitative tests near the patient. Besides the reading instrument where the data are read, the spinit®, there is a set of multi-parametric disposable disks, that through the use of a simple drop of blood and without the need of using additional reagents by the user, makes possible to provide quantitative results in less than 15 minutes. One of the developed assays was the C-reactive protein, whose disposable hold several detection zones which include a biological recognition layer, using antibody fragments immobilized on a gold surface. When the patient’s blood plasma passes through this zones, the target marker binds to the immobilized antibody fragments. This phenomenon induces a change in the refractive index which subsequently is amplified through the binding with gold nanoparticles. The underlying technology of this reader is the surface plasmon ressonance. In this context, this dissertation had as initial goal the assay time reduction without changing the detection ranges for C-reactive protein. Throughout this study it was verified that with the systems’ existent gold nanoparticles size it wouldn’t be possible to fulfill the objective, since the detection range was diminished. This was achieved using 20 nm gold nanoparticles that allowed, beside assay time reduction (main objective), to improve the detection range. The results obtained allowed to analytically validate the product’s improvement and to submit a European Conformity marking, since the C-reactive protein detection range was improved from 5 to 100 mg/L to 4 to 180 mg/L and the assay time shortened from 15 to 6 minutes.
Point-of-care technology emerged in order to allow a quick and efficient diagnosis, making possible a timely intervention and treatment by healthcare professionals. Biosurfit SA. is a company that developed and produced a medical equipment that allows to perform several quantitative tests near the patient. Besides the reading instrument where the data are read, the spinit®, there is a set of multi-parametric disposable disks, that through the use of a simple drop of blood and without the need of using additional reagents by the user, makes possible to provide quantitative results in less than 15 minutes. One of the developed assays was the C-reactive protein, whose disposable hold several detection zones which include a biological recognition layer, using antibody fragments immobilized on a gold surface. When the patient’s blood plasma passes through this zones, the target marker binds to the immobilized antibody fragments. This phenomenon induces a change in the refractive index which subsequently is amplified through the binding with gold nanoparticles. The underlying technology of this reader is the surface plasmon ressonance. In this context, this dissertation had as initial goal the assay time reduction without changing the detection ranges for C-reactive protein. Throughout this study it was verified that with the systems’ existent gold nanoparticles size it wouldn’t be possible to fulfill the objective, since the detection range was diminished. This was achieved using 20 nm gold nanoparticles that allowed, beside assay time reduction (main objective), to improve the detection range. The results obtained allowed to analytically validate the product’s improvement and to submit a European Conformity marking, since the C-reactive protein detection range was improved from 5 to 100 mg/L to 4 to 180 mg/L and the assay time shortened from 15 to 6 minutes.
Descrição
Tese de mestrado, Biologia Molecular e Genética, Universidade de Lisboa, Faculdade de Ciências, 2016
Palavras-chave
Tecnologia point-of-care Spinit Biosurfit Ressonância de plasmão de superfície Proteína C reativa Teses de mestrado - 2016