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Journal of Visualized Experiments
Article . 2024 . Peer-reviewed
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Assay Development for High-Throughput Drug Screening Against Mycobacteria

Authors: Oliveira, Gabriel S.; Bento, Clara M.; Pombinho, António; Reis, Rita; Van Calster, Kevin; De Vooght, Linda; Maia, André F.; +3 Authors

Assay Development for High-Throughput Drug Screening Against Mycobacteria

Abstract

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.

Keywords

Mycobacterium abscessus / drug effects, Mycobacterium abscessus, Pharmacology. Therapy, Mycobacterium Infections, Nontuberculous / microbiology, Drug Evaluation, Preclinical, Mycobacterium Infections, Nontuberculous, Anti-Bacterial Agents / pharmacology, Mycobacterium Infections, Nontuberculous / drug therapy, High-Throughput Screening Assays, Anti-Bacterial Agents, High-Throughput Screening Assays / methods, Drug Evaluation, Preclinical / methods, Anti-Bacterial Agents / chemistry, Humans

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selected citations
These citations are derived from selected sources.
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
0
Average
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