Surface-based detection of nucleic acid in a convection flow fluidic device

Abstract

The present disclosure provides methods, composition and devices for performing convection-based PCR and non-enzymatic amplification of nucleic acid sequences. Techniques and reagents employed in these methods include toehold probes, strand displacement reactions, Rayleigh-Benard convection, temperature gradients, multiplexed amplification, multiplexed detection, and DNA functionalization, in openand closed systems, for use in nucleic tests and assays.

Description

[0001] priority statement [0002] This application claims the benefit of priority to US Provisional Application Serial No. 62 / 314,909, filed March 29, 2016, which is incorporated herein by reference in its entirety. technical field [0003] The present disclosure describes novel reagents, apparatus and methods for the detection and quantification of specific nucleic acid sequences for scientific and clinical research and diagnostic applications. Background technique [0004] Most commercial nucleic acid (NA) assays require the use of enzymes for molecular or signal amplification. Enzymes such as DNA polymerases have been optimized to be rapid and specific. Reconstitution of lyophilized enzymes in resource-limited conditions reduces the need for cold chains. Isothermal nucleic acid amplification assays such as NEAR, LAMP, and NASBA allow DNA / RNA mapping without complex temperature cycling equipment. Despite many of these advances, existing nucleic acid detection technolo...

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Problems solved by technology

In these systems, there are false positive amplifications due to DNA "breathing" events, which result in the release of amplicon molecules in the absence of the detected target sequence

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