Droplet microfluidic platform for fast and continuous-flow RT-qPCR analysis devoted to cancer diagnosis application

RT-qPCR represents a key method in cancer diagnostic, however the constant increase in patients and cancer biomarker panels to screen requires the implementation of faster approaches allowing smaller reagents and samples volumes consumption. To fulfil these needs, we present here a fully automated droplet microfluidics platform that couples a specifically designed thermal system with a fluorescence excitation/detection module. Additionally, the droplet generation and merging approaches allows the analysis of multiple samples and genes with no risk of contamination. This platform has been initially validated by investigating HER2 overexpression in two cancer cell lines starting from total RNA samples; these results have been successfully compared with those obtained by a commercially available machine, showing no limitation in terms of number of processed samples as well as reducing of 200 times the analysis costs per patient. Finally, we have demonstrated its capability in performing fast RT-qPCR, raising the throughput of analysis to a hundred samples in less than 20 min.