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Phenotypic drug susceptibility testing for Mycobacterium tuberculosis variant bovis BCG in 12 hours
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Cell and Molecular Biology, Molecular Systems Biology. Uppsala University, Science for Life Laboratory, SciLifeLab.ORCID iD: 0000-0002-3440-8974
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Cell and Molecular Biology, Molecular Systems Biology. Uppsala University, Science for Life Laboratory, SciLifeLab.ORCID iD: 0000-0001-7355-9751
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Medical Cell Biology. Uppsala University, Science for Life Laboratory, SciLifeLab.ORCID iD: 0009-0009-6487-5803
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Cell and Molecular Biology, Molecular Systems Biology. Uppsala University, Science for Life Laboratory, SciLifeLab.ORCID iD: 0000-0002-5925-8669
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2025 (English)In: Nature Communications, E-ISSN 2041-1723, Vol. 16, no 1, article id 4366Article in journal (Refereed) Published
Abstract [en]

Drug-resistant tuberculosis (DR-TB) kills similar to 200,000 people every year. A contributing factor is the slow turnaround time (TAT) associated with drug susceptibility diagnostics. The prevailing gold standard for phenotypic drug susceptibility testing (pDST) takes at least two weeks. Here we show that growth-based pDST for slow-growing mycobacteria can be conducted in 12 h. We use Mycobacterium tuberculosis variant bovis Bacillus Calmette-Guerin (BCG) and Mycobacterium smegmatis as the mycobacterial pathogen models and expose them to antibiotics used in (multidrug-resistant) tuberculosis (TB) treatment regimens - i.e., rifampicin (RIF), isoniazid (INH), ethambutol (EMB), linezolid (LZD), streptomycin (STR), bedaquiline (BDQ), and levofloxacin (LFX). The bacterial growth in a microfluidic chip is tracked by time-lapse phase-contrast microscopy. A deep neural network-based segmentation algorithm is used to quantify the growth rate and to determine how the strains responded to drug treatments. Most importantly, a panel of susceptible and resistant M. bovis BCG are tested at critical concentrations for INH, RIF, STR, and LFX. The susceptible strains could be identified in less than 12 h. These findings are comparable to what we expect for pathogenic M. tuberculosis as they share 99.96% genetic identity.

Place, publisher, year, edition, pages
Springer Nature, 2025. Vol. 16, no 1, article id 4366
National Category
Infectious Medicine Microbiology in the Medical Area
Identifiers
URN: urn:nbn:se:uu:diva-559309DOI: 10.1038/s41467-025-59736-9ISI: 001501680700004PubMedID: 40348759Scopus ID: 2-s2.0-105005235872OAI: oai:DiVA.org:uu-559309DiVA, id: diva2:1971390
Funder
Swedish Foundation for Strategic Research, SSF ARC19-0016Knut and Alice Wallenberg Foundation, 2023.0531Novo Nordisk, 0083419Available from: 2025-06-17 Created: 2025-06-17 Last updated: 2025-06-17Bibliographically approved

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Tran, Buu MinhLarsson, JimmyGrip, AnastasiaKarempudi, PraneethElf, Johan

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