Abstracts

This presentation will focus on the design and development of bioluminescence-based point-of-care diagnostics methods. Specifically, we will discuss laboratory methods, as well as point-of-care (POC) testing that have been developed by us based on the bioluminescence detection principle. These diagnostic methods are designed for monitoring infectious diseases. Examples of POC methods employed to address an unmet need regarding the diagnosis of urinary tract infection (UTI) - antimicrobial resistance (AMR) and COVID-19 will be discussed. Emphasis will be on the simple design principles and improving the performance/stability of bioluminescent proteins to enhance their utility in POC method development. Rapid bioluminescence assays were developed and evaluated for the detection of UTI using intact lyophilized cells of Photobacterium leiognathi ATCC 33981™. Further, using ATP-based assay antimicrobial resistance was detected. UTI-AMR detection was performed using a microtiter-based, hand-held luminometer, and POC adaptor-cartridge-based detection. To improve the usage of bioluminescent reporters in POC devices, we prepared an encapsulated, stable bioluminescent fusion protein, Tamavidin-Gaussia luciferase (TA2-Gluc) in a ZIF-8 metal-organic framework (MOF). Tamavidin-2 is a group of avidin-like high-affinity biotin-binding protein and Gluc is a small Renilla-type luciferase that yields high bioluminescent intensity. This bioluminescent reporter produces light upon the addition of the substrate coelenterazine. We evaluated the stability of the ZIF-8 encapsulated TA2-GLuc at various temperatures (25° C, 4° C, and room temperature) and demonstrated that we can preserve this fusion protein in all of the tested temperatures for at least 6 months. Further, we designed a highly sensitive bioluminescent assay for detecting the SARS-CoV-2 spike antigen using ZIF-8 encapsulated TA2-Gluc.