Dispositivos microfluídicos baseados em fita adesiva dupla face e aplicações no diagnóstico molecular da Covid-19

• Ano de defesa: 2021
• Autor(a) principal: Estrela, Paulo Felipe Neves
• Orientador(a): Duarte, Gabriela Rodrigues Mendes
• Banca de defesa: Duarte, Gabriela Rodrigues Mendes, Gomes, Danielle Cangussu de Castro, Rodrigues, Flávia Melo
• Tipo de documento: Dissertação
• Tipo de acesso: Acesso aberto
• Idioma: por
• Instituição de defesa: Universidade Federal de Goiás
• Programa de Pós-Graduação: Programa de Pós-graduação em Química (IQ)
• Departamento: Instituto de Química - IQ (RG)
• País: Brasil
• Palavras-chave em Português: Microchip; SARS-CoV-2; Bioanalítica
• Palavras-chave em Inglês: Loop-mediated isothermal amplification; Bioanalytics
• Área do conhecimento CNPq: CIENCIAS EXATAS E DA TERRA::QUIMICA::QUIMICA-ANALITICA
• Link de acesso: http://repositorio.bc.ufg.br/tede/handle/tede/11612

Microfluidic devices have shown potential application in several areas of science due to the possibility of integration and automation of steps, reducing time and cost in total analysis. However, due to the laboriousness of the preparation, its use is still discreet. Thus, in the aim to make microfluidic devices accessible, in this study we explore the development of disposable microfluidic devices based on double-sided tape and polyester film (DF-Pe) and we initially demonstrated its use in all stages of molecular diagnosis for COVID-19. Devices based on double-sided adhesive tape were manufactured in two different forms: using a laser cutter or totally free-instrumentation. The DF-Pe microchips developed here exhibited low cost, simplicity, fast fabrication, chemical resistance, versatility for layout changes and biocompatibility. In the DF-Pe microchips, we carried out the isolation of nucleic acids from samples of patients with SARS-CoV-2 infection, using the technique of dynamic solid phase extraction (dSPE) with an efficiency of extraction of 56%, obtaining the RNA at the end of the process suitable to be amplified by RT-qPCR (Reverse Transcription - Quantitative Pol-ymerase Chain Reaction) and RT-LAMP (Reverse Transcription - Loop-Mediated Isother-mal Amplification). We demonstrated a RT-LAMP based assay to detect SARS-CoV-2 di-rectly on microdevices in 30 minutes, also we applied the test to a panel of 40 samples and the results obtained were with 100% concordance with the gold standard method (RT-qPCR). Furthermore, the features of the DF-Pe microdevice were evaluated through the development of passive micromixers that allowed visual detection of the amplification prod-ucts directly on the microdevices. The high simplicity, cost-effective and fast manufacturing of the DF-Pe microdevices, integrated to the simple and quick extraction, amplification and viral detection directly on chip, demonstrates the high potential application of this method in the molecular diagnosis of COVID-19 in resources limited settings.