Abstract
Bioluminescent signaling systems derived from marine microorganisms provide sensitive, self-amplified readouts that enable rapid diagnostic testing without complex laboratory instrumentation. This dissertation describes the development and clinical evaluation of bioluminescence-based platforms for point-of-care detection of urinary tract infections (UTIs), antimicrobial resistance (AMR), and infectious disease biomarkers. UTIs are among the most common bacterial infections worldwide, and rising AMR, combined with delays in conventional diagnostics, contributes to inappropriate antimicrobial use. Two whole-cell bioluminescent assays, TuBETUr (Tube Bioluminescence Extinction Technology Urine) and CUBET (Cellphone-based UTI Bioluminescence Extinction Technology), were developed using viable or lyophilized Photobacterium leiognathi and detected clinically relevant uropathogens, including Escherichia coli, Proteus mirabilis, Staphylococcus aureus, and Candida albicans, at concentrations below the diagnostic threshold of 10⁵ CFU/mL in spiked and clinical urine samples. TuBETUr correctly identified all culture-positive specimens (n = 29), while CUBET enabled statistically significant smartphone-based differentiation between UTI-positive and negative samples (p ≤ 0.0001). Building on these platforms, a tandem bioluminescent assay enabled UTI detection within five minutes at bacterial concentrations as low as 2 × 10⁴ CFU/mL and AMR profiling within 30 minutes using a modified ATP-based assay, with no interference from urinary components; in a clinical pilot study (n = 8), the assay demonstrated 100% concordance with standard methods. Additionally, a ZIF-8–stabilized tamavidin2–Gaussia luciferase reporter enabled room-temperature storage and sensitive ELISA-based detection of SARS-CoV-2 antigens, supporting the clinical potential of bioluminescent diagnostics to improve timely infection detection, guide antimicrobial therapy, and expand access to point-of-care testing in resource-limited settings.