journal article·2025-07-31Open access

Mycobacteriophages in the Treatment of Mycobacterial Infections: From Compassionate Use to Targeted Therapy

Magdalena Druszczynska, Beata Sadowska, Agnieszka Zablotni, Lesia Zhuravska, Jakub Kulesza, Marek Fol

Applied Sciences· Volume 15· Issue 15

Abstract

This review addresses the urgent need for alternative strategies to combat drug-resistant mycobacterial infections, including multidrug-resistant (MDR) and extensively drug-resistant (XDR) tuberculosis, as well as non-tuberculous mycobacterial (NTM) diseases. Traditional antibiotics are increasingly limited by resistance, toxicity, and poor efficacy, particularly in immunocompromised patients. A comprehensive literature search was conducted using PubMed, Scopus, and Google Scholar, covering publications primarily from 2000 to 2025. Only articles published in English were included to ensure consistency in data interpretation. Search terms included “mycobacteriophages,” “phage therapy,” “drug-resistant mycobacteria, “diagnostic phages,” and “phage engineering.” The review examines the therapeutic and diagnostic potential of mycobacteriophages—viruses that specifically infect mycobacteria—focusing on their molecular biology, engineering advances, delivery systems, and clinical applications. Evidence suggests that mycobacteriophages offer high specificity, potent bactericidal activity, and adaptability, positioning them as promising candidates for targeted therapy. Although significant obstacles remain—including immune interactions, limited host range, and regulatory challenges—rapid progress in synthetic biology and delivery platforms continues to expand their clinical potential. As research advances and clinical frameworks evolve, mycobacteriophages are poised to become a valuable asset in the fight against drug-resistant mycobacterial diseases, offering new precision-based solutions where conventional therapies fail.
Source: crossref · Quality: baseline

Research context

OpenAlex topics: Bacteriophages and microbial interactions · Monoclonal and Polyclonal Antibodies Research · Mycobacterium research and diagnosis

SciVX discovery tags are terminology-based. OpenAlex topics are source-provided classifications.

Affiliations

Department of Immunology and Infectious Biology, Institute of Microbiology, Biotechnology and Immunology, Faculty of Biology and Environmental Protection, University of Lodz, 90-237 Lodz, Poland

Department of Bacterial Biology, Institute of Microbiology, Biotechnology and Immunology, Faculty of Biology and Environmental Protection, University of Lodz, 90-237 Lodz, Poland

Department of Internal Diseases and Clinical Pharmacology, Medical University of Lodz, 90-237 Lodz, Poland

References (100+)

Show source-provided references
1. (2025, July 18). Global Tuberculosis Report 2024. Available online: https://www.who.int/publications/i/item/9789240101531.
2. Multidrug-resistant and extensively drug-resistant tuberculosis: Consequences for the global HIV community10.1097/QCO.0b013e3283210020 ↗
3. Exploring diagnostic methods for drug-resistant tuberculosis: A comprehensive overview10.1016/j.tube.2024.102522 ↗
4. Ototoxicity among multidrug-resistant TB patients: A systematic review and meta-analysis10.5588/ijtld.20.0217 ↗
5. Rates and risk factors for nephrotoxicity and ototoxicity among tuberculosis patients in Tbilisi, Georgia10.5588/ijtld.18.0626 ↗
6. Actionable mechanisms of drug tolerance and resistance in Mycobacterium tuberculosis10.1111/febs.17142 ↗
7. Multicenter, retrospective cohort study of antimycobacterial treatment-related harms among patients with non-tuberculosis Mycobacterium infections in the United States10.1128/aac.01596-24 ↗
8. Conyers, L.E., and Saunders, B.M. (2024). Treatment for non-tuberculous mycobacteria: Challenges and prospects. Front. Microbiol., 15.10.3389/fmicb.2024.1394220 ↗
9. Gu, Y., Nie, W., Huang, H., and Yu, X. (2023). Non-tuberculous mycobacterial disease: Progress and advances in the development of novel candidate and repurposed drugs. Front. Cell Infect. Microbiol., 13.10.3389/fcimb.2023.1243457 ↗
10. Phage therapy as a renewed therapeutic approach to mycobacterial infections: A comprehensive review10.2147/IDR.S218638 ↗
11. Opperman, C.J., and Brink, A.J. (2025). Phage Therapy for Mycobacteria: Overcoming Challenges, Unleashing Potential. Infect. Dis. Rep., 17.10.3390/idr17020024 ↗
12. Ouyang, X., Li, X., Song, J., Wang, H., Wang, S., Fang, R., Li, Z., and Song, N. (2023). Mycobacteriophages in diagnosis and alternative treatment of mycobacterial infections. Front. Microbiol., 14.10.3389/fmicb.2023.1277178 ↗
13. Yang, F., Labani-Motlagh, A., Bohorquez, J.A., Moreira, J.D., Ansari, D., Patel, S., Spagnolo, F., Florence, J., Vankayalapati, A., and Sakai, T. (2024). Bacteriophage therapy for the treatment of Mycobacterium tuberculosis infections in humanized mice. Commun. Biol., 7.10.1038/s42003-024-06006-x ↗
14. Shield, C.G., Swift, B.M.C., McHugh, T.D., Dedrick, R.M., Hatfull, G.F., and Satta, G. (2021). Application of Bacteriophages for Mycobacterial Infections, from Diagnosis to Treatment. Microorganisms, 9.10.3390/microorganisms9112366 ↗
15. Bacteriophages and their genomes10.1016/j.coviro.2011.06.009 ↗
16. Dark Matter of the Biosphere: The Amazing World of Bacteriophage Diversity10.1128/JVI.01340-15 ↗
17. Understanding the enormous diversity of bacteriophages: The tailed phages that infect the bacterial family Enterobacteriaceae10.1016/j.virol.2014.08.024 ↗
18. Sun, X., Jiang, H., and Zhang, S. (2024). Diversities and interactions of phages and bacteria in deep-sea sediments as revealed by metagenomics. Front. Microbiol., 14.10.3389/fmicb.2023.1337146 ↗
19. Filik, K., Szermer-Olearnik, B., Oleksy, S., Brykała, J., and Brzozowska, E. (2022). Bacteriophage Tail Proteins as a Tool for Bacterial Pathogen Recognition—A Literature Review. Antibiotics, 11.10.3390/antibiotics11050555 ↗
20. Bacteriophage prehistory: Is or is not Hankin, 1896, a phage reference?10.4161/bact.1.3.16591 ↗
21. Bacteriophage Therapy: Exploiting Smaller Fleas10.1086/597405 ↗
22. Phage Therapy—Everything Old is New Again10.1155/2006/329465 ↗
23. Bacteriophage taxonomy10.1071/MA11090 ↗
24. Diallo, K., and Dublanchet, A. (2023). A Century of Clinical Use of Phages: A Literature Review. Antibiotics, 12.10.3390/antibiotics12040751 ↗
25. Wei, J., Peng, N., Liang, Y., Li, K., and Li, Y. (2020). Phage Therapy: Consider the Past, Embrace the Future. Appl. Sci., 10.10.3390/app10217654 ↗
26. Ferry, T., Kolenda, C., Briot, T., Souche, A., Lustig, S., Josse, J., Batailler, C., Pirot, F., Medina, M., and Leboucher, G. (2021). Past and Future of Phage Therapy and Phage-Derived Proteins in Patients with Bone and Joint Infection. Viruses, 13.10.3390/v13122414 ↗
27. Keen, E.C. (2012). Phage therapy: Concept to cure. Front. Microbiol., 3.10.3389/fmicb.2012.00238 ↗
28. Bacteriophage Taxonomy: A Continually Evolving Disci-pline10.1007/978-1-0716-3523-0_3 ↗
29. Elois, M.A., Silva, R.D., Pilati, G.V.T., Rodríguez-Lázaro, D., and Fongaro, G. (2023). Bacteriophages as Biotechnological Tools. Viruses, 15.10.3390/v15020349 ↗
30. How to introduce a new bacteriophage on the block: A short guide to phage classification10.1128/jvi.01821-23 ↗
31. Structure and function of bacteriophage T410.2217/fmb.14.91 ↗
32. Bacteriophage lambda: Early pioneer and still relevant10.1016/j.virol.2015.02.010 ↗
33. Bacteriophage deoxyribonucleate infection of competent Bacillus subtilis10.1128/jb.89.3.782-790.1965 ↗
34. Characterization of the DNA-protein complex at the termini of the bacteriophage PRD1 genome10.1128/jvi.50.2.309-315.1984 ↗
35. Properties of bacteriophage PM2: A lipid-containing bacterial virus10.1016/0042-6822(68)90094-9 ↗
36. Mycoplasmatales virus-laidlawii 2, a new virus isolated from Acholeplasma laidlawii10.1099/0022-1317-12-1-65 ↗
37. Gottlieb, P., and Alimova, A. (2023). Discovery and Classification of the φ6 Bacteriophage: An Historical Review. Viruses, 15.10.3390/v15061308 ↗
38. Structure and assembly of filamentous bacteriophages10.1016/j.pbiomolbio.2014.02.003 ↗
39. Bacteriophages and phage-inspired nanocarriers for targeted delivery of therapeutic cargos10.1016/j.addr.2016.03.003 ↗
40. Gardner, G.M., and Weiser, R.S. (1947). A bacteriophage for Mycobacterium smegmatis. Proc. Soc. Exp. Biol. Med., 66.10.3181/00379727-66-16037 ↗
41. Killing of Mycobacterium avium and Mycobacterium tuberculosis by a Mycobacteriophage Delivered by a Nonvirulent Mycobacterium: A Model for Phage Therapy of Intracellular Bacterial Pathogens10.1086/343812 ↗
42. Mycobacteriophages: Genes and genomes10.1146/annurev.micro.112408.134233 ↗
43. Comparative Genomic Analysis of 60 Mycobacteriophage Genomes: Genome Clustering, Gene Acquisition, and Gene Size10.1016/j.jmb.2010.01.011 ↗
44. Comparative genomics of the mycobacteriophages: Insights into bacteriophage evolution10.1016/j.resmic.2008.04.008 ↗
45. Abolishment of morphology-based taxa and change to binomial species names: 2022 taxonomy update of the ICTV bacterial viruses subcommittee10.1007/s00705-022-05694-2 ↗
46. Turner, D., Kropinski, A.M., and Adriaenssens, E.M. (2021). A Roadmap for Genome-Based Phage Taxonomy. Viruses, 13.10.3390/v13030506 ↗
47. Practical methods for determining phage growth parameters10.1007/978-1-60327-164-6_18 ↗
48. Phage classification and characterization10.1007/978-1-60327-164-6_13 ↗
49. Science Education Alliance Phage Hunters Advancing Genomics and Evolutionary Science Program; KwaZulu-Natal Research Institute for Tuberculosis and HIV Mycobacterial Genetics Course Students; Phage Hunters Integrating Research and Education Program. Complete genome sequences of 138 mycobacteriophages10.1128/JVI.06870-11 ↗
50. Hatfull, G.F. (2018). Mycobacteriophages. Microbiol. Spectr., 6.10.1128/microbiolspec.GPP3-0026-2018 ↗
51. Rees, C., and Botsaris, G. (2012). The Use of Phage for Detection, Antibiotic Sensitivity Testing and Enumeration [Internet]. Understanding Tuberculosis—Global Experiences and Innovative Approaches to the Diagnosis, InTech.10.5772/29734 ↗
52. Weiss, S.M., Happy, K.K., Baliraine, F.W., Beach, A.K., Brobston, S.M., Martinez, C.P., Menard, K.J., Orton, S.M., Salazar, A.L., and Frederick, G.D. (2023). Complete genome sequences and characteristics of seven novel mycobacteriophages isolated in East Texas. Microbiol. Resour. Announc., 12.10.1128/mra.00335-23 ↗
53. Diversity among the tailed-bacteriophages that infect the Enterobacteriaceae10.1016/j.resmic.2008.04.005 ↗
54. Genome structure of mycobacteriophage D29: Implications for phage evolution10.1006/jmbi.1997.1610 ↗
55. Mycobacteriophage TM4: Genome structure and gene expression10.1054/tuld.1998.0007 ↗
56. On the nature of mycobacteriophage diversity and host preference10.1016/j.virol.2012.09.026 ↗
57. Molecular Genetics of Mycobacteriophages10.1128/microbiolspec.MGM2-0032-2013 ↗
58. (2025, June 23). The Actinobacteriophage Database. Available online: https://phagesdb.org/.
59. Phage Therapy for Nontuberculous Mycobacteria: Challenges and Opportunities10.1007/s41030-022-00210-y ↗
60. Hatfull, G.F. (2014). Mycobacteriophages: Windows into tuberculosis. PLoS Pathog., 10.10.1371/journal.ppat.1003953 ↗
61. Engineered bacteriophages for treatment of a patient with a disseminated drug-resistant Mycobacterium abscessus10.1038/s41591-019-0437-z ↗
62. Mycobacterial nucleoid-associated protein Lsr2 is required for productive mycobacteriophage infection10.1038/s41564-023-01333-x ↗
63. Khambhati, K., Bhattacharjee, G., Gohil, N., Dhanoa, G.K., Sagona, A.P., Mani, I., Bui, N.L., Chu, D.T., Karapurkar, J.K., and Jang, S.H. (2022). Phage engineering and phage-assisted CRISPR-Cas delivery to combat multi-drug-resistant pathogens. Bioeng. Transl. Med., 8.10.1002/btm2.10381 ↗
64. Zein-Eddine, R., Refrégier, G., Cervantes, J., and Yokobori, N.K. (2023). The future of CRISPR in Mycobacterium tuberculosis infection. J. Biomed. Sci., 30.10.1186/s12929-023-00932-4 ↗
65. CRISPR-mediated phage resistance and the ghost of coevolution past
66. SEA-PHAGES and SEA-GENES: Advancing Virology and Science Education10.1146/annurev-virology-113023-110757 ↗
67. Whole genome comparison of a large collection of mycobacteriophages reveals a continuum of phage genetic diversity10.7554/eLife.06416 ↗
68. Actinobacteriophages: Genomics, Dynamics, and Applications10.1146/annurev-virology-122019-070009 ↗
69. Phage therapy: Awakening a sleeping giant10.1042/ETLS20170002 ↗
70. Phage Therapy of Mycobacterium Infections: Compassionate Use of Phages in 20 Patients With Drug-Resistant Mycobacterial Disease10.1093/cid/ciac453 ↗
71. Recent advances in bacteriophage therapy: How delivery routes, formulation, concentration and timing influence the success of phage therapy10.1111/j.2042-7158.2011.01324.x ↗
72. Al-Ishaq, R.K., Skariah, S., and Büsselberg, D. (2020). Bacteriophage Treatment: Critical Evaluation of Its Application on World Health Organi-zation Priority Pathogens. Viruses, 13.10.3390/v13010051 ↗
73. Phage treatment of human infections10.4161/bact.1.2.15845 ↗
74. The role of regulated clinical trials in the development of bacteriophage therapeutics10.4172/1747-0862.1000050 ↗
75. Oral Phage Therapy of Acute Bacterial Diarrhea with Two Coliphage Preparations: A Randomized Trial in Children From Bangladesh10.1016/j.ebiom.2015.12.023 ↗
76. The phage therapy paradigm: Prêt-à-porter or sur-mesure?10.1007/s11095-010-0313-5 ↗
77. Podlacha, M., Grabowski, Ł., Kosznik-Kawśnicka, K., Zdrojewska, K., Stasiłojć, M., Węgrzyn, G., and Węgrzyn, A. (2021). Interactions of Bacterio-phages with Animal and Human Organisms-Safety Issues in the Light of Phage Therapy. Int. J. Mol. Sci., 22.10.3390/ijms22168937 ↗
78. Pros and cons of phage therapy10.4161/bact.1.2.14590 ↗
79. Merabishvili, M., Pirnay, J.P., Verbeken, G., Chanishvili, N., Tediashvili, M., Lashkhi, N., Glonti, T., Krylov, V., Mast, J., and Van Parys, L. (2009). Quali-ty-controlled small-scale production of a well-defined bacteriophage cocktail for use in human clinical trials. PLoS ONE, 4.10.1371/journal.pone.0004944 ↗
80. Phages limit the evolution of bacterial antibiotic resistance in experimental microcosms10.1111/j.1752-4571.2011.00236.x ↗
81. Hitchcock, N.M., Devequi Gomes Nunes, D., Shiach, J., Valeria Saraiva Hodel, K., Dantas Viana Barbosa, J., Alencar Pereira Rodrigues, L., Coler, B.S., Botelho Pereira Soares, M., and Badaró, R. (2023). Current Clinical Landscape and Global Potential of Bacteriophage Therapy. Viruses, 15.10.3390/v15041020 ↗
82. The next generation of bacteriophage therapy10.1016/j.mib.2011.07.028 ↗
83. A century of bacteriophage research and applications: Impacts on biotechnology, health, ecology and the economy10.1002/jctb.5810 ↗
84. Bacteriophages as therapeutic agents10.3109/07853890108995959 ↗
85. Żaczek, M., Weber-Dąbrowska, B., Międzybrodzki, R., Łusiak-Szelachowska, M., and Górski, A. (2020). Phage Therapy in Poland—A Centennial Journey to the First Ethically Approved Treatment Facility in Europe. Front. Microbiol., 11.10.3389/fmicb.2020.01056 ↗
86. Bacteriophage therapy of bacterial infections: An update of our institute’s experience
87. Nontuberculous mycobacterial infections: A clinical review10.1007/s15010-004-4001-4 ↗
88. Wade, M.M. (2004). Mechanisms of drug resistance in Mycobacterium Tuberculosis. Front. Biosci., 9.10.2741/1289 ↗
89. Evolution of Drug-Resistant Tuberculosis: A Tale of Two Species10.1073/pnas.91.7.2428 ↗
90. Role of the Clinical Mycobacteriology Laboratory in Diagnosis and Management of Tuberculosis in Low-Prevalence Settings10.1128/JCM.02451-10 ↗
91. Detection of tuberculosis (TB) using gold standard method, direct sputum smears microscopy, PCR, qPCR and electrochemical DNA sensor: A mini review10.54987/jobimb.v4i2.305 ↗
92. Bartolomeu-Gonçalves, G., de Souza, J.M., Fernandes, B.T., Spoladori, L.F.A., Correia, G.F., de Castro, I.M., Borges, P.H.G., Silva-Rodrigues, G., Tavares, E.R., and Yamauchi, L.M. (2024). Tuberculosis Diagnosis: Current, Ongoing, and Future Approaches. Diseases, 12.10.3390/diseases12090202 ↗
93. Nalugwa, T., Shete, P.B., Nantale, M., Farr, K., Ojok, C., Ochom, E., Mugabe, F., Joloba, M., Dowdy, D.W., and Moore, D.A.J. (2020). Challenges with Scale-up of GeneXpert MTB/RIF® in Uganda: A Health Systems Perspective. BMC Health Serv. Res., 20.10.1186/s12913-020-4997-x ↗
94. Fighting Persistence: How Chronic Infections with Mycobacterium Tuberculosis Evade T Cell-Mediated Clearance and New Strategies to Defeat Them10.1128/IAI.00916-19 ↗
95. Phage as a Diagnostic—The Use of Phage in TB Diagnosis10.1002/jctb.439 ↗
96. Phage-Based Platforms for the Clinical Detection of Human Bacterial Pathogens10.4161/bact.19274 ↗
97. Reporter phage and breath tests: Emerging phenotypic assays for diagnosing active tuberculosis, antibiotic resistance, and treatment efficacy10.1093/infdis/jir454 ↗
98. Rapid Film-Based Determination of Antibiotic Susceptibilities of Mycobacterium Tuberculosis Strains by Using a Luciferase Reporter Phage and the Bronx Box10.1128/JCM.37.4.1144-1149.1999 ↗
99. Luciferase Reporter Phage (LRP) Assay for Anti Tuberculosis Screening: Current Status and Challenges10.21786/bbrc/13.3/39 ↗
100. Luciferase Reporter Mycobacteriophages for Detection, Identification, and Antibiotic Susceptibility Testing of Mycobacterium tuberculosis in Mexico10.1128/JCM.39.11.3883-3888.2001 ↗