A self-contained diagnostic platform for DNA concentration, elution, and qPCR inside a LabCard with stored reagents

摘要

This paper presents a self contained system consisting of a portable platform and a disposable LabCard capable of performing a nucleic acid concentration, amplification, and detection. The LabCard comprises several inlets and outlets, integrated microvalves, two chambers and stored reagents. In the first chamber it is performed a magnetic beads-based DNA concentration, washing and elution. Then, the eluted DNA is transferred to the second chamber where it rehydrates with the pre-stored reagents. Finally, a real-time Polymerase Chain Reaction (qPCR) takes place and the fluorescence signal is recorded and displayed in real time. The magnetic bead protocol allows the LabCard to process a wide range of sample volumes (from 10 ul to 10 ml). The PCR reagents are stored inside the LabCard using a gelification technique allowing: (i) long-term storage of the LabCards, (ii) reduction of the reagent volumes, and (iii) significant simplification of the LabCard design and user operation. The cross contamination is avoided by making the LabCard disposable. On the other hand, the platform has been conceived as a comprehensive and generic solution able to perform a wide range of biological assays. These assays consist in an automated and programmable sequence of sample preparation phases followed by a DNA amplification and detection phase.