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Publicação
Fast prototyping microfluidics: Integrating droplet digital lamp for absolute quantification of cancer biomarkers
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Orientador(es)
Resumo(s)
Microfluidic (MF) advancements have been leveraged toward the development of state-of-the-art platforms for molecular diagnostics, where isothermal amplification schemes allow for further simplification of DNA detection and quantification protocols. The MF integration with loop-mediated isothermal amplification (LAMP) is today the focus of a new generation of chip-based devices for molecular detection, aiming at fast and automated nucleic acid analysis. Here, we combined MF with droplet digital LAMP (ddLAMP) on an all-in-one device that allows for droplet generation, target amplification, and absolute quantification. This multilayer 3D chip was developed in less than 30 minutes by using a low-cost and extremely adaptable production process that exploits direct laser writing technology in “Shrinky-dinks” polystyrene sheets. ddLAMP and target quantification were performed directly on-chip, showing a high correlation between target concentration and positive droplet score. We validated this integrated chip via the amplification of targets ranging from five to 500,000 copies/reaction. Furthermore, on-chip amplification was performed in a 10 µL volume, attaining a limit of detection of five copies/µL under 60 min. This technology was applied to quantify a cancer biomarker, c-MYC, but it can be further extended to any other disease biomarker.
Descrição
UID/CTM/50025/2019 UID/Multi/04378/2019 Inn-INDIGO/0002/2015 PTDC/BTM-SAL/31201/2017 SFRH/BPD/124311/2016
Palavras-chave
Digital amplification Integrated microfluidics device Lab-on-a-chip Loop-mediated isothermal amplification Oncogenes Analytical Chemistry Biochemistry Atomic and Molecular Physics, and Optics Instrumentation Electrical and Electronic Engineering SDG 3 - Good Health and Well-being SDG 9 - Industry, Innovation, and Infrastructure