Desenvolvimento de kits para rápido diagnóstico da dengue utilizando nanoparticulas de ouro
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Data
2019-02-22
Autores
Basso, Caroline Rodrigues
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Universidade Estadual Paulista (Unesp)
Resumo
Doenças tropicais são aquelas causadas por agentes infecciosos ou parasitas e consideradas endêmicas em populações na sua maioria de baixa renda, residente em países tropicais e subtropicais. Dentre essas doenças temos a dengue, que acomete milhares de pessoas levando a mortes todos os anos. A dengue é transmitida através da picada do mosquito contaminado e a movimentação populacional a torna de fácil disseminação em todo o território nacional. Atualmente, os exames para diagnóstico desta doença são realizados em laboratórios, principalmente testes sorológicos. Porém, esta metodologia apresenta como limitação o custo elevado além dos resultados levarem dias para que fiquem prontos. Por isso, se faz necessário o desenvolvimento de uma nova metodologia confiável e de baixo custo para um rápido diagnóstico. Atualmente, nanobiossensores vem se destacando como uma ferramenta analítica que pode identificar a presença de patógenos utilizando como base nanoestruturas. Logo, o objetivo do presente projeto foi desenvolver e padronizar uma nova metodologia para o diagnóstico rápido e eficaz do vírus da dengue, utilizando diferentes nanoestruturas modificadas com a deposição de anticorpos específicos e aptâmeros. As metodologias foram comparadas mediante as técnicas de ressonância plasmônica de superfície localizada, ressonância plasmônica de superfície, microscopia eletrônica de transmissão, espectroscopia de impedância eletroquímica, espalhamento dinâmico de luz e análise do software ImageJ. Os resultados obtidos com as técnicas apresentaram algumas vantagens em relação aos métodos tradicionais de detecção, como rápido tempo de análise e baixo custo, mostrando ser altamente eficaz na detecção dos quatro sorotipos da dengue, com uma abordagem simples, levando em média 30 minutos para obtenção dos resultados. Mudanças colorimétricas nas etapas de modificações utilizando nanopartículas maghemita, proporcionaram rápida detecção visual do vírus sem o uso de equipamento, tornando possível o diagnóstico em locais distantes dos laboratórios. O valor final do kit foi de cerca de R$ 9,00 por amostra.
Tropical diseases are those caused by infectious agents or parasites and are considered endemic in mostly low-income populations residing in tropical and subtropical countries. Among these diseases we have the dengue that causes thousands of deaths each year. Dengue is transmitted through the bite of the contaminated mosquito and the population movement makes it easily spread throughout the national territory. Currently, the diagnostic test of this disease is performed in laboratories, being serological tests. However this methodology presents as a limitation the higher cost and the delay for the result come out. Therefore, it is necessary to develop a new, reliable and low-cost methodology for rapid diagnosis. Nowadays, nanobiosensors have been highlighted with analytical tool that can identify the presence of pathogens using as base nanostructures. The objective of the present project was to develop and standardize a new methodology for a fast and effective diagnosis of dengue virus using using different nanostructures modified with the deposition of specific antibodies and aptamers. The methodology was compared by localized surface plasmon resonance, surface plasmon resonance, transmission electron microscopy, electrochemical impedance spectroscopy, dynamic light scattering techniques and ImageJ software analysis. The results obtained with the techniques presented some advantages over the traditional methods of detection, such as fast analysis and low cost, showing to be highly effective in the detection of the four dengue serotypes with a simple approach, taking an average of 30 minutes to obtain the results. Colorimetric changes in the steps of modifications using maghemite nanoparticles provided fast visual detection of the virus without the use of equipment, making it possible diagnosis in remote locations of the laboratories. The final value of the kit was about R$ 9.00 per sample.
Tropical diseases are those caused by infectious agents or parasites and are considered endemic in mostly low-income populations residing in tropical and subtropical countries. Among these diseases we have the dengue that causes thousands of deaths each year. Dengue is transmitted through the bite of the contaminated mosquito and the population movement makes it easily spread throughout the national territory. Currently, the diagnostic test of this disease is performed in laboratories, being serological tests. However this methodology presents as a limitation the higher cost and the delay for the result come out. Therefore, it is necessary to develop a new, reliable and low-cost methodology for rapid diagnosis. Nowadays, nanobiosensors have been highlighted with analytical tool that can identify the presence of pathogens using as base nanostructures. The objective of the present project was to develop and standardize a new methodology for a fast and effective diagnosis of dengue virus using using different nanostructures modified with the deposition of specific antibodies and aptamers. The methodology was compared by localized surface plasmon resonance, surface plasmon resonance, transmission electron microscopy, electrochemical impedance spectroscopy, dynamic light scattering techniques and ImageJ software analysis. The results obtained with the techniques presented some advantages over the traditional methods of detection, such as fast analysis and low cost, showing to be highly effective in the detection of the four dengue serotypes with a simple approach, taking an average of 30 minutes to obtain the results. Colorimetric changes in the steps of modifications using maghemite nanoparticles provided fast visual detection of the virus without the use of equipment, making it possible diagnosis in remote locations of the laboratories. The final value of the kit was about R$ 9.00 per sample.
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Palavras-chave
biossensores, dengue, nanopartículas, biosensors, nanoparticles