Nanoimunoensaios para diagnóstico de malária
Resumo
O desenvolvimento de Testes de Diagnóstico Rápido (TDR) para o diagnóstico de malária é uma área em intenso desenvolvimento. Aqui, descrevemos dois nanoimunoensaios para a deteção do parasita da malária; um baseado em eletroforese em gel de agarose; e outro utilizando a tecnologia lab-on-paper. Ambos os nanoimunoensaios usam nanopartículas de ouro (AuNP) funcionalizadas com o ácido 11-mercaptoundecanóico (MUA) ou com o pentapéptido CALNN, conjugadas com o anticorpo monoclonal (anti-PfHRP2) que reconhece especificamente o antigénio de Plasmodium falciparum Histidine Rich Protein 2 (PfHRP2). A formação dos bionanoconjugados AuNP-anti-PfHRP2 foi comprovada por eletroforese em gel de agarose, demonstrando que a funcionalização da AuNP e o modo de conjugação (por fisiossorção ou por reticulação química) controlam a ligação do anticorpo à AuNP. O nanoimunoensaio baseado em gel de agarose, mostrou que os bionanoconjugados AuNP-MUA-anti-PfHRP2 detetaram uma concentração de antigénio de 12 μg/mL, uma ordem de grandeza inferior à detetada pelos bionanoconjugados AuNP-CALNN-anti-PfHRP2 (700 μg/mL). Os bionanoconjugados AuNP-MUA-anti-PfHRP2 também revelaram ser eficazes para a deteção colorimétrica direta do antigénio por Western Blot, eliminando a necessidade de um anticorpo secundário. O TDR baseado na tecnologia lab-on-paper foi desenvolvido numa tira de papel de filtro à base de celulose, eliminando o tradicional invólucro de plástico. A quantidade de bionanoconjugados e os componentes das linhas de teste e de controlo foram otimizados. Futuramente, o nosso objetivo é aplicar este TDR na análise de amostras clínicas.
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