Aufdembrink, Lauren2025-03-212025-03-212022-12https://hdl.handle.net/11299/270573University of Minnesota Ph.D. dissertation. December 2022. Major: Molecular, Cellular, Developmental Biology and Genetics. Advisor: Aaron Engelhart. 1 computer file (PDF); xiii, 193 pages + 2 supplementary files.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.enFluorescence monitoringFluorescent aptamersIsothermal nucleic acid amplificationpathogen detectionportable reaction monitoringThesis or Dissertation