Assay Development for High-Throughput Drug Screening Against Mycobacteria
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| Titel: | Assay Development for High-Throughput Drug Screening Against Mycobacteria |
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| Autoren: | Oliveira, Gabriel S., Bento, Clara M., Pombinho, António, Reis, Rita, Van Calster, Kevin, De Vooght, Linda, Maia, André F., Cos, Paul, Gomes, Maria Salomé, Silva, Tânia |
| Weitere Verfasser: | Instituto de Investigação e Inovação em Saúde |
| Quelle: | Journal of visualized experiments |
| Verlagsinformationen: | MyJove Corporation, 2024. |
| Publikationsjahr: | 2024 |
| Schlagwörter: | 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 |
| Beschreibung: | 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. |
| Publikationsart: | Article |
| Dateibeschreibung: | application/pdf |
| Sprache: | English |
| ISSN: | 1940-087X |
| DOI: | 10.3791/66860 |
| Zugangs-URL: | https://pubmed.ncbi.nlm.nih.gov/39526621 https://hdl.handle.net/10216/165538 https://hdl.handle.net/10067/2093020151162165141 https://repository.uantwerpen.be/docstore/d:irua:25946 |
| Dokumentencode: | edsair.doi.dedup.....951dca4a98885b6013c64a335a36eaab |
| Datenbank: | OpenAIRE |
Nájsť tento článok vo Web of Science | 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. |
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| ISSN: | 1940087X |
| DOI: | 10.3791/66860 |