Detection technologies employing fluorescent aptamers

Abstract

Isothermal, cell-free, synthetic biology-based approaches to pathogen detection leverage the power of tools available in biological systems, such as highly active polymerases compatible with lyophilization, without the complexity inherent to live-cell systems. Among cell-free techniques, nucleic acid sequence based amplification (NASBA) is well known. Another synthetic biology tool for quantification and visualization is fluorescent aptamers, which are nucleic acid-base, GFP-like tags. This dissertation describes work in which I impart a fluorescent real-time readout on NASBA employing fluorescent aptamers, called Apta-NASBA. This generates an inexpensive, field deployable, single-pot pathogen detection reaction. Additionally, a durable, low-cost, and portable fluorescence detection platform capable of monitoring Apta-NASBA reactions is engineered with a 3D-printed chassis and easily obtainable components. Apta-NASBA and the fluorescence detection platform allow for affordable pathogen detection compatible with field diagnostics and use in low resource areas.