Citation:

Oliveira, B, Veigas B, Fernandes AR, Aguas H, Martins R, Fortunato E, Baptista PV.  2020.  Fast Prototyping Microfluidics: Integrating Droplet Digital Lamp for Absolute Quantification of Cancer Biomarkers, 2020. Sensors (Basel). 20(6)

Abstract:

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 microL volume, attaining a limit of detection of five copies/microL 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.

Notes:

Oliveira, BeatrizVeigas, Bruno
Fernandes, Alexandra R
Aguas, Hugo
Martins, Rodrigo
Fortunato, Elvira
Baptista, Pedro Viana
eng
UID/CTM/50025/2019; UID/Multi/04378/2019; Inn-INDIGO/0002/2015; PTDC/BTM-SAL/31201/2017; SFRH/BPD/124311/2016/Fundacao para a Ciencia e Tecnologia
Switzerland
Sensors (Basel). 2020 Mar 14;20(6). pii: s20061624. doi: 10.3390/s20061624.

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