| dc.description.abstract | Tuberculosis, the disease caused by the mycobacterial pathogen Mycobacterium tuberculosis, has been, prior to the COVID-19 pandemic, the world’s deadliest infectious disease. The illness has been afflicting humans for centuries, and despite decades long efforts to eradicate it and the availability of curative treatment options, tuberculosis remains prevalent across most of the globe. According to the WHO Tuberculosis Reports, approximately 1.7 billion people are estimated to be infected by the bacteria and 1.5 million people die from the disease every year. Effective diagnostics and treatment are key to combating any infectious disease pandemic and in this work, we present a novel approach to improving both. We created a magnetophoretic microfluidic device which uses recombinant mycobacteriophage tail fiber proteins bound to magnetic nanoparticles to pull down mycobacterial cells, selectively concentrating the cells before performing a diagnostic microbiological stain protocol. We have shown that we could lower the limit of detection of Mycobacterium tuberculosis from a synthetic sputum sample by 6 to 26-folds per milliliter of sample, without significantly altering the process used at point-of-care clinics. We also investigated the synergistic interactions between drugs, which we believe is a key element to improving efficacy of drug regimen, but also for the creation of new drug combinations, specifically designed to work together. We have shown that using low doses of Bedaquiline causes inhibitors of PEPCK to be synthetically lethal, despite their lack of whole cell activity on their own. This suggests that typical drug discovery campaign may be missing some valuable compounds, that could play an important part of a combination regimen. Furthermore, we have synthesized the necessary substrate required for the development of an enzymatic assay for peptidyl tRNA hydrolase. This assay could then be used to identify inhibitors of PTH, which could restore Mtb sensitivity to macrolides. These drug discovery campaigns seek to make a better use of synergistic drug interaction and use this information as an integral part of drug discovery. | |
