| Autor: |
Oliveira GS; i3S - Instituto de Investigação e Inovação em Saúde da Universidade do Porto; ICBAS - Instituto de Ciências Biomédicas Abel Salazar, Universidade do Porto., Bento CM; i3S - Instituto de Investigação e Inovação em Saúde da Universidade do Porto; IBMC - Instituto de Biologia Molecular e Celular, Universidade do Porto; Programa Doutoral em Biologia Molecular e Celular (MCBiology), Instituto de Ciências Biomédicas Abel Salazar da Universidade do Porto., Pombinho A; i3S - Instituto de Investigação e Inovação em Saúde da Universidade do Porto; IBMC - Instituto de Biologia Molecular e Celular, Universidade do Porto., Reis R; i3S - Instituto de Investigação e Inovação em Saúde da Universidade do Porto; IBMC - Instituto de Biologia Molecular e Celular, Universidade do Porto., Van Calster K; Laboratory for Microbiology, Parasitology and Hygiene (LMPH); Department of Infectious Diseases in Humans, Immune Response, Sciensano., De Vooght L; Laboratory for Microbiology, Parasitology and Hygiene (LMPH)., Maia AF; i3S - Instituto de Investigação e Inovação em Saúde da Universidade do Porto; IBMC - Instituto de Biologia Molecular e Celular, Universidade do Porto., Cos P; Laboratory for Microbiology, Parasitology and Hygiene (LMPH)., Gomes MS; i3S - Instituto de Investigação e Inovação em Saúde da Universidade do Porto; ICBAS - Instituto de Ciências Biomédicas Abel Salazar, Universidade do Porto., Silva T; i3S - Instituto de Investigação e Inovação em Saúde da Universidade do Porto; ICBAS - Instituto de Ciências Biomédicas Abel Salazar, Universidade do Porto; tania.silva@i3s.up.pt. |
| Abstrakt: |
Mycobacterium abscessus (Mab) infections are challenging to treat due to high intrinsic drug resistance, comparable to multidrug-resistant tuberculosis. Treatments are extremely ineffective and based on a multi-drug regimen, resulting in low patient compliance. Consequently, the scientific community is urged to identify new and effective drugs to treat these infections. One of the strategies employed to this end is drug repurposing - the process of identifying new therapeutic opportunities for existing drugs in the market, circumventing the time required to establish pharmacokinetic and safety profiles of new drugs. With most studies on drug development against Mab relying on traditional and time-consuming methods, an assay for high-throughput drug screening was developed against mycobacteria using an in house developed double-reporter strain of Mab. Using liquid-handling robotics, automated microscopy, and analysis, alongside in house developed double reporter strains, bacterial viability can be rapidly measured using two different readouts, luminescence and fluorescence, without adding reagents or performing any extra steps. This reduces time and variability between assays, a major advantage for high-throughput screenings. The described protocol was validated by screening a library of 1280 compounds. The obtained results were corroborated by the literature, with efficient detection of active compounds. Thus, this work fulfilled the aim of supplying the field with a new tool to help fight this extremely drug-resistant bacteria. |
