Novel paper-microfluidic analytical device (uPADs) coupled with Phage-amplification and Quantum dots for multiplexed pathogen diagnostics.

Current methods of species-specific bacterial detection and identification are complex, time-consuming, and often require expensive specialized equipment and highly trained personnel. There is a growing need for developing fieldable devices to combat both naturally occurring, and deliberate outbreaks of biothreat agents. Building upon Guild BioSciences previous research on phage-mediated pathogen detection and lateral flow paper-microfluidic devices (uPADs), we propose to develop a fieldable pathogen diagnostic tool that synergistically combines the advantages of bacteriophages, fluorescent quantum dots (QDS) with the simplicity and portability of uPADs. The proposed device will have the capability to rapidly detect and identify multiple biodefense relevant pathogens in a field setting. Specifically, in the phase I project, we will: 1) generate affinity-tagged bacteriophages, 2) design and fabricate uPADs, 3) demonstrate the multiplexed and sensitive detection of B. anthracis and Y. pestis by phage-amplification assay via incorporating QDs as signal readout, 4) determine the assay sensitivity and limit-of-detection (LoD) in simulated clinical samples, and 5) validate the uPADs detection with in-house developed bioluminescent-phage-based pathogen detection. Phase II will optimize the assay process and reagents, generate the preclinical data required for FDA-approval, design a prototype fieldable kit, and expand the technology to enable the detection of additional bacterial pathogens.