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Assay Development for High-Throughput Drug Screening Against Mycobacteria.

Gabriel S Oliveira1, Clara M Bento2, António Pombinho3

  • 1i3S - Instituto de Investigação e Inovação em Saúde da Universidade do Porto; ICBAS - Instituto de Ciências Biomédicas Abel Salazar, Universidade do Porto.

Journal of Visualized Experiments : Jove
|November 11, 2024
PubMed
Summary

Developing new treatments for Mycobacterium abscessus (Mab) infections is crucial due to high drug resistance. A novel high-throughput screening assay was developed to rapidly identify effective drugs against this challenging bacteria.

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Area of Science:

  • Microbiology
  • Drug Discovery
  • Infectious Diseases

Background:

  • Mycobacterium abscessus infections present significant treatment challenges due to intrinsic drug resistance, similar to multidrug-resistant tuberculosis.
  • Current treatment regimens are often ineffective, leading to poor patient compliance and a critical need for novel therapeutic strategies.
  • Drug repurposing offers a promising avenue for identifying new treatments by leveraging existing drugs, thus expediting the drug development process.

Purpose of the Study:

  • To develop and validate a high-throughput drug screening assay for identifying novel therapeutic agents against Mycobacterium abscessus.
  • To establish a rapid and reliable method for assessing bacterial viability in drug screening, reducing time and variability.
  • To facilitate the discovery of effective drugs to combat drug-resistant Mycobacterium abscessus infections.

Main Methods:

  • Development of an in-house double-reporter strain of Mycobacterium abscessus for enhanced screening capabilities.
  • Utilization of liquid-handling robotics, automated microscopy, and analysis for rapid bacterial viability measurement via luminescence and fluorescence readouts.
  • Validation of the assay by screening a library of 1280 compounds to assess its efficacy in detecting active agents.

Main Results:

  • The developed assay enables rapid measurement of bacterial viability using dual luminescence and fluorescence readouts without additional reagents or steps.
  • Screening of a 1280-compound library successfully identified active compounds, with results corroborated by existing literature.
  • The assay demonstrated efficiency in detecting compounds active against Mycobacterium abscessus, confirming its utility.

Conclusions:

  • A novel, efficient high-throughput screening assay has been successfully developed and validated for Mycobacterium abscessus drug discovery.
  • This tool significantly reduces assay time and variability, offering a valuable resource for the scientific community in the fight against drug-resistant infections.
  • The assay provides a much-needed platform to accelerate the identification of new drugs to combat challenging Mycobacterium abscessus infections.