ArticleThe Journal of infectious diseases2025
Engineered Mycobacteriophage TM4::GeNL Rapidly Determines Bedaquiline, Pretomanid, Linezolid, Rifampicin, and Clofazimine Sensitivity in Mycobacterium tuberculosis Clinical Isolates.
Article in The Journal of infectious diseases, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 6 papers.
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Who cites it
6 citing papers in PubMed.
- Rapid detection of treatment-emergent resistance to bedaquiline and pretomanid inMicrobiology spectrum · 2026Article
- Rapid drug susceptibility testing of Mycobacterium tuberculosis against first-line drugs simultaneously all-in-one plate by using a novel high-sensitive reporter phage combined with the BACTEC MGIT 960 system.Infectious diseases of poverty · 2026Article
- Advances in mycobacteriophage research: from lytic mechanisms to one health applications.Frontiers in cellular and infection microbiology · 2026Review
- Luciferase reporter mycobacteriophage (TM4::Journal of clinical microbiology · 2025Article
- Mycobacteriophage-mediated gene transfer enables in vitro drug screening and in vivo tracking ofProceedings of the National Academy of Sciences of the United States of America · 2025Article
- Rapid detection of drug resistance inFrontiers in cellular and infection microbiology · 2025Article
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Authors and funding
19 authors.
Funding
Abstract
backgroundDrug-resistant tuberculosis is a growing public health threat, and early characterization of the resistance phenotype is essential for guiding treatment and mitigating the high mortality associated with the disease. However, the slow growth rate of Mycobacterium tuberculosis, the causative agent of tuberculosis, necessitates several weeks for conventional culture-dependent drug susceptibility testing (DST). In addition, there are no widely available molecular diagnostic assays for evaluating resistance to newer tuberculosis drugs or drugs with complex resistance mechanisms.
methodsWe have developed a luciferase-based reporter mycobacteriophage assay that can determine drug resistance within 48 hours. We engineered the TM4 mycobacteriophage to express green enhanced nanoluciferase (GeNL) cassette and optimized DST for bedaquiline, pretomanid, linezolid, clofazimine, and rifampicin using clinical M. tuberculosis isolates.
resultsTo assess the feasibility of this assay, we conducted a proof-of-principle study using 53 clinical M. tuberculosis isolates. TM4::GeNL phage DST effectively distinguished between sensitive and resistant isolates for bedaquiline and rifampicin at a concentration of 0.125 μg/mL. Optimal differentiation between sensitive and resistant isolates for pretomanid, clofazimine, and linezolid was achieved at concentrations of 0.5 μg/mL, 0.25 μg/mL, and 1 μg/mL, respectively. Additionally, TM4::GeNL DST identified low-level rifampicin resistance in clinical isolates even though they were classified as sensitive by Mycobacteria Growth Indicator Tube DST.
conclusionsTM4::GeNL reporter phage DST offers a rapid method to identify M. tuberculosis drug resistance, including resistance to newer tuberculosis drugs.
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