@article {16155, title = {Fast Prototyping Microfluidics: Integrating Droplet Digital Lamp for Absolute Quantification of Cancer Biomarkers}, journal = {Sensors (Basel)}, volume = {20}, year = {2020}, note = {

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.

}, month = {2020}, 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.

}, keywords = {*Biosensing Techniques, Biomarkers, Tumor/genetics/*isolation \& purification, digital amplification, DNA, Neoplasm/genetics/*isolation \& purification, Humans, integrated microfluidics device, lab-on-a-chip, Lab-On-A-Chip Devices, Limit of Detection, loop-mediated isothermal amplification, Microfluidics/methods, Molecular Diagnostic Techniques/methods, Neoplasms/*diagnosis/genetics/pathology, Nucleic Acid Amplification Techniques/methods, oncogenes, Pathology, Molecular/methods}, isbn = {1424-8220 (Electronic)1424-8220 (Linking) }, url = {https://www.ncbi.nlm.nih.gov/pubmed/32183359}, author = {Oliveira, B. and B. Veigas and Fernandes, A. R. and Aguas, H. and R. Martins and E. Fortunato and P.V. Baptista} }