Phenotypic drug susceptibility testing for Mycobacterium tuberculosis variant bovis BCG in 12 hours

Abstract Drug-resistant tuberculosis (DR-TB) kills ~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 sho...

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Main Authors: Buu Minh Tran, Jimmy Larsson, Anastasia Grip, Praneeth Karempudi, Johan Elf
Format: Article
Language:English
Published: Nature Portfolio 2025-05-01
Series:Nature Communications
Online Access:https://doi.org/10.1038/s41467-025-59736-9
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author Buu Minh Tran
Jimmy Larsson
Anastasia Grip
Praneeth Karempudi
Johan Elf
author_facet Buu Minh Tran
Jimmy Larsson
Anastasia Grip
Praneeth Karempudi
Johan Elf
author_sort Buu Minh Tran
collection DOAJ
description Abstract Drug-resistant tuberculosis (DR-TB) kills ~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-Guérin (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.
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spelling doaj-art-108dbf85c17248b986581bf8b2b2cabc2025-08-20T03:53:13ZengNature PortfolioNature Communications2041-17232025-05-0116111210.1038/s41467-025-59736-9Phenotypic drug susceptibility testing for Mycobacterium tuberculosis variant bovis BCG in 12 hoursBuu Minh Tran0Jimmy Larsson1Anastasia Grip2Praneeth Karempudi3Johan Elf4Department of Cell and Molecular Biology, SciLifeLab, Uppsala UniversityDepartment of Cell and Molecular Biology, SciLifeLab, Uppsala UniversityDepartment of Cell and Molecular Biology, SciLifeLab, Uppsala UniversityDepartment of Cell and Molecular Biology, SciLifeLab, Uppsala UniversityDepartment of Cell and Molecular Biology, SciLifeLab, Uppsala UniversityAbstract Drug-resistant tuberculosis (DR-TB) kills ~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-Guérin (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.https://doi.org/10.1038/s41467-025-59736-9
spellingShingle Buu Minh Tran
Jimmy Larsson
Anastasia Grip
Praneeth Karempudi
Johan Elf
Phenotypic drug susceptibility testing for Mycobacterium tuberculosis variant bovis BCG in 12 hours
Nature Communications
title Phenotypic drug susceptibility testing for Mycobacterium tuberculosis variant bovis BCG in 12 hours
title_full Phenotypic drug susceptibility testing for Mycobacterium tuberculosis variant bovis BCG in 12 hours
title_fullStr Phenotypic drug susceptibility testing for Mycobacterium tuberculosis variant bovis BCG in 12 hours
title_full_unstemmed Phenotypic drug susceptibility testing for Mycobacterium tuberculosis variant bovis BCG in 12 hours
title_short Phenotypic drug susceptibility testing for Mycobacterium tuberculosis variant bovis BCG in 12 hours
title_sort phenotypic drug susceptibility testing for mycobacterium tuberculosis variant bovis bcg in 12 hours
url https://doi.org/10.1038/s41467-025-59736-9
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