antibioticsАнтибиотики и ХимиотерапияAntibiot Khimioter = Antibiotics and Chemotherapy0235-2990ООО «Издательство ОКИ»10.37489/0235-2990-2025-70-11-12-63-74WRGKMLantibiotics-1313Research ArticleОБЗОРЫREVIEWSНативные и модифицированные микобактериальные бактериофаги: источники, технологии получения и применение в диагностике и терапии микобактериозовNative and Engineered Mycobacteriophages: Sources, Manufacturing Technologies, and Applications in Diagnosis and Therapy of Mycobacterioseshttps://orcid.org/0000-0003-0536-8149АбдрахмановаР. О.AbdrakhmanovaR. O.

Абдрахманова Радмила Охасовна — ассистент кафедры микробиологии и вирусологии, научный сотрудник Научно-исследовательского центра ФГБОУ ВО ГМУ Минздрава России.

Астрахань

Radmila O. Abdrakhmanova — Assistant of the Department of Microbiology and Virology, Researcher of the Research Center, Astrakhan State Medical University of the Ministry of Health of Russian Federation.

Astrakhan

https://orcid.org/0000-0002-2904-9276РубальскийО. В.RubalskyO. V.

Рубальский Олег Васильевич — д. м. н., профессор, заведующий кафедрой микробиологии и вирусологии, главный научный сотрудник Научно-исследовательского центра ФГБОУ ВО «Астраханский ГМУ» Минздрава России.

Астрахань

Oleg V. Rubalsky — D. Sc. in Medicine, Professor, Head of the Department of Microbiology and Virology, Chief Researcher at the Research Center, Astrakhan State Medical University of the Ministry of Health of the Russian Federation.

Astrakhan

https://orcid.org/0000-0001-6245-2553ЛазькоМ. В.LazkoM. V.

Лазько Марина Владимировна — д. б. н., профессор, начальник управления науки, профессор кафедры микробиологии и вирусологии, заведующая лабораторией Научно-исследовательского центра ФГБОУ ВО Астраханский ГМУ Минздрава России.

Астрахань

Marina V. Lazko — D. Sc. in Biology, Professor, Head of the Science Department, Professor of the Department of Microbiology and Virology, Head of the Laboratory of the Research Center, Astrakhan State Medical University.

Astrakhan

https://orcid.org/0000-0003-2998-2864ЯсенявскаяА. Л.YasenyavskayaA. L.

Ясенявская Анна Леонидовна — д. м. н., доцент, руководитель Научно-исследовательского центра, профессор кафедры фармакогнозии, фармацевтической технологии и биотехнологии, ФГБОУ ВО «Астраханский ГМУ» Минздрава России.

Астрахань

Anna L. Yasenyavskaya — D. Sc. in Medicine, Associate Professor, Head of the Research Center, Professor at the Department of Pharmacognosy, Pharmaceutical Technology, and Biotechnology, Astrakhan State Medical University of the Ministry of Health of the Russian Federation.

Astrakhan

yasen_9@mail.ruhttps://orcid.org/0000-0003-0838-7353РубальскаяТ. С.RubalskaiaT. S.

Рубальская Татьяна Сергеевна — консультант ООО «ГорДиаМед».

Астрахань

Tatiana S. Rubalskaia — consultant GorDiaMed LLC.

Astrakhan

Астраханский государственный медицинский университетРоссияAstrakhan State Medical UniversityRussian FederationООО «ГорДиаМед»РоссияGorDiaMed LLCRussian Federation2025040320267011-126374Copyright © Абдрахманова Р.О., Рубальский О.В., Лазько М.В., Ясенявская А.Л., Рубальская Т.С., 20262026Абдрахманова Р.О., Рубальский О.В., Лазько М.В., Ясенявская А.Л., Рубальская Т.С.Abdrakhmanova R.O., Rubalsky O.V., Lazko M.V., Yasenyavskaya A.L., Rubalskaia T.S.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.antibiotics-chemotherapy.ru/jour/article/view/1313

Микобактериальные бактериофаги (микобактериофаги) — вирусы, инфицирующие представителей рода Mycobacterium, рассматриваются как быстро развивающаяся платформа для диагностики и терапии инфекций, вызванных туберкулёзными и нетуберкулезными микобактериями (НТМ). В обзоре критически обобщены современные подходы к поиску и выделению нативных микобактериофагов из природных субстратов; биологической и морфологической характеристике микобактериофагов; их молекулярной идентификации и аннотации геномов; технологиям направленной модификации и созданию рекомбинантных фагов микобактерий с заданными свойствами; требованиям к разработке лекарственных форм с микобактериальными фагами, контролю качества и биобезопасности этих форм; стратегиям персонализированной фаготерапии НТМ-инфекций и потенциалу диагностических репортерных фагов при микобактериозах. Обсуждаются ограничения (узкий спектр литической активности, возможная резистентность бактерий, взаимодействие с иммунной системой), регуляторные и технологические барьеры, а также перспективы интеграции микобактериофаговых платформ с антибиотикотерапией и биоинженерией. Предложены стандартизованные рабочие схемы пробоподготовки, скрининга, очистки и контроля качества препаратов, содержащих микобактериофаги, а также минимальные наборы исследовательской и клинической документации.

Mycobacteriophages – viruses that infect members of the genus Mycobacterium — are emerging as a rapidly advancing platform for the diagnosis and treatment of infections caused by both tuberculous and nontuberculous mycobacteria (NTM). This review critically summarizes current approaches to (i) the discovery and isolation of native mycobacteriophages from environmental substrates; (ii) their biological and morphological characterization; (iii) molecular identification and genome annotation; (iv) targeted modification and construction of recombinant phages (including BRED) with desired properties; (v) development of pharmaceutical formulations, quality control, and biosafety; and (vi) strategies for personalized phage therapy of NTM infections, as well as the potential of reporter phages for diagnostics. Limitations (narrow host range activity, emergence of bacterial resistance, interactions with the immune system), regulatory and technological barriers, and perspectives on integrating phage platforms with antibiotic therapy and bioengineering are discussed. Standardized workflows for sampling, screening, purification, and quality control, as well as minimal research and clinical documentation sets, are proposed.

микобактериофагнетуберкулёзные микобактериифаготерапиярекомбинантные фагирепортерные фагиконтроль качестваперсонализированная медицинаmycobacteriophagenontuberculous mycobacteriaphage therapyrecombinant phagesreporter phagesquality controlpersonalized medicineРабота выполнена в рамках государственного задания Министерства здравоохранения Российской Федерации в части проведения прикладной НИР по теме «Разработка тест-систем для скрининговой ПЦР-диагностики, мониторинга течения микобактериозов и видовой ПЦР-идентификации микобактерий».ReferencesWorld Health Organization. Global tuberculosis report 2024. Geneva: WHO; 2024.World Health Organization. Global tuberculosis report 2024. Geneva: WHO; 2024.Estaji F., Kamali A., Keikha M. Strengthening the global response to tuberculosis: insights from the 2024 WHO global TB report. J Clin Tuberc Other Mycobact Dis. 2025; 39: 100522. doi: 10.1016/j.jctube.2025.100522.Estaji F., Kamali A., Keikha M. Strengthening the global response to tuberculosis: insights from the 2024 WHO global TB report. J Clin Tuberc Other Mycobact Dis. 2025; 39: 100522. doi: 10.1016/j.jctube.2025.100522.Murray C. J. L., Ikuta K. S., Sharara F. et al. Global burden of bacterial antimicrobial resistance in 2019: a systematic analysis. Lancet. 2022; 399 (10325): 629–655. doi: 10.1016/S0140-6736(21)02724-0.Murray C. J. L., Ikuta K. S., Sharara F. et al. Global burden of bacterial antimicrobial resistance in 2019: a systematic analysis. Lancet. 2022; 399 (10325): 629–655. doi: 10.1016/S0140-6736(21)02724-0.Prevots D. R., Marras J. E., Wagner D., Morimoto K. Global epidemiology of nontuberculous mycobacterial pulmonary disease: a review. Clin Chest Med. 2023; 44 (4): 675–721. doi: 10.1016/j.ccm.2023.08.012.Prevots D. R., Marras J. E., Wagner D., Morimoto K. Global epidemiology of nontuberculous mycobacterial pulmonary disease: a review. Clin Chest Med. 2023; 44 (4): 675–721. doi: 10.1016/j.ccm.2023.08.012.Harada K. et al. Trends in nontuberculous mycobacterial disease mortality worldwide, 2000-2022. Int J Infect Dis. 2025; [Epub ahead of print].Harada K. et al. Trends in nontuberculous mycobacterial disease mortality worldwide, 2000-2022. Int J Infect Dis. 2025; [Epub ahead of print].Daley C. L., Iaccarino J. M., Lange C., Cambau E., Wallace Jr. R. J., Andrejak C. et al. Treatment of nontuberculous mycobacterial pulmonary disease: an official ATS/ERS/ESCMID/IDSA guideline. Clin Infect Dis. 2020; 71 (4): e1-e36. doi: 10.1093/cid/ciaa241.Daley C. L., Iaccarino J. M., Lange C., Cambau E., Wallace Jr. R. J., Andrejak C. et al. Treatment of nontuberculous mycobacterial pulmonary disease: an official ATS/ERS/ESCMID/IDSA guideline. Clin Infect Dis. 2020; 71 (4): e1-e36. doi: 10.1093/cid/ciaa241.Dedrick R. M., Guerrero-Bustamante C., Garlena R. A., Russell D., Ford K., Harris K. et al. Engineered bacteriophages for treatment of a patient with a disseminated drug-resistant Mycobacterium abscessus infection. Nat Med. 2019; 25 (5): 730–733. doi: 10.1038/s41591-019-0437-z.Dedrick R. M., Guerrero-Bustamante C., Garlena R. A., Russell D., Ford K., Harris K. et al. Engineered bacteriophages for treatment of a patient with a disseminated drug-resistant Mycobacterium abscessus infection. Nat Med. 2019; 25 (5): 730–733. doi: 10.1038/s41591-019-0437-z.Dedrick R. M., Smith B. E., Cristinziano M., Freeman K. G., Jacobs-Sera D., Belessis Y. et al. Phage therapy of Mycobacterium infections: compassionate use of phages in 20 patients with drug-resistant mycobacterial disease. Clin Infect Dis. 2023; 76 (1): 103–112. doi: 10.1093/cid/ciac453.Dedrick R. M., Smith B. E., Cristinziano M., Freeman K. G., Jacobs-Sera D., Belessis Y. et al. Phage therapy of Mycobacterium infections: compassionate use of phages in 20 patients with drug-resistant mycobacterial disease. Clin Infect Dis. 2023; 76 (1): 103–112. doi: 10.1093/cid/ciac453.Banaiee N., Bobadilla-del-Valle M., Riska P. F., Small P. M., Ponce-De-Leon A., Jacobs Jr. W. R. et al. Luciferase reporter mycobacteriophages for detection, identification, and antibiotic susceptibility testing of Mycobacterium tuberculosis. J Clin Microbiol. 2001; 39 (11): 3883–3888. doi: 10.1128/JCM.39.11.3883-3888.2001.Banaiee N., Bobadilla-del-Valle M., Riska P. F., Small P. M., Ponce-De-Leon A., Jacobs Jr. W. R. et al. Luciferase reporter mycobacteriophages for detection, identification, and antibiotic susceptibility testing of Mycobacterium tuberculosis. J Clin Microbiol. 2001; 39 (11): 3883–3888. doi: 10.1128/JCM.39.11.3883-3888.2001.Mayer O., Jain P., Weisbrod T. R., Biro D., Ho L., Jacobs-Sera D. et al. Fluorescent reporter DS6A mycobacteriophages reveal unique variations in infectibility and phage production in mycobacteria. J Bacteriol. 2016; 198 (23): 3220–3232. doi: 10.1128/JB.00592-16.Mayer O., Jain P., Weisbrod T. R., Biro D., Ho L., Jacobs-Sera D. et al. Fluorescent reporter DS6A mycobacteriophages reveal unique variations in infectibility and phage production in mycobacteria. J Bacteriol. 2016; 198 (23): 3220–3232. doi: 10.1128/JB.00592-16.Kropinski A. M., Mazzocco A., Waddell T. E., Lingohr E., Johnson R. P. Enumeration of bacteriophages by double agar overlay plaque assay. Methods Mol Biol. 2009; 501: 69–76. doi: 10.1007/978-1-60327-164-6_7.Kropinski A. M., Mazzocco A., Waddell T. E., Lingohr E., Johnson R. P. Enumeration of bacteriophages by double agar overlay plaque assay. Methods Mol Biol. 2009; 501: 69–76. doi: 10.1007/978-1-60327-164-6_7.Clokie M. R. J., Kropinski A. M. eds. Bacteriophages: Methods and Protocols, Volume 1: Isolation, Characterization, and Interactions. New York: Humana Press; 2009.Clokie M. R. J., Kropinski A. M. eds. Bacteriophages: Methods and Protocols, Volume 1: Isolation, Characterization, and Interactions. New York: Humana Press; 2009.Göller P. C., Haro-Moreno J. M., Rodriguez-Valera F., Loessner M. J., Gomez-Sanz E. Uncovering a hidden diversity: optimized protocols for extraction of dsDNA bacteriophages from soil. Sci Rep. 2020; 8 (1): 17. doi: 10.1186/s40168-020-0795-2.Göller P. C., Haro-Moreno J. M., Rodriguez-Valera F., Loessner M. J., Gomez-Sanz E. Uncovering a hidden diversity: optimized protocols for extraction of dsDNA bacteriophages from soil. Sci Rep. 2020; 8 (1): 17. doi: 10.1186/s40168-020-0795-2.World Health Organization. Laboratory biosafety manual, 4th ed. Geneva: WHO; 2020.World Health Organization. Laboratory biosafety manual, 4th ed. Geneva: WHO; 2020.CDC/NIH. Biosafety in Microbiological and Biomedical Laboratories (BMBL), 6th ed. 2020.CDC/NIH. Biosafety in Microbiological and Biomedical Laboratories (BMBL), 6th ed. 2020.Monpoeho S., Maul A., Mignotte-Cadiergues B., Schwartzbrrod L., Billaudel S., Ferre V. Best viral elution method for enteroviruses from sludge. Appl Environ Microbiol. 2001; 67 (6): 2484–2488. doi: 10.1128/AEM.67.6.2484-2488.2001.Monpoeho S., Maul A., Mignotte-Cadiergues B., Schwartzbrrod L., Billaudel S., Ferre V. Best viral elution method for enteroviruses from sludge. Appl Environ Microbiol. 2001; 67 (6): 2484–2488. doi: 10.1128/AEM.67.6.2484-2488.2001.Hill V. R., Narayanan J., Gallen R. R., Ferdinand K., Cromeans T., Vinje J. Simultaneous recovery of DNA/RNA from diverse microbes. Pathogens. 2015; 4 (2): 335–351. doi: 10.3390/pathogens4020335.Hill V. R., Narayanan J., Gallen R. R., Ferdinand K., Cromeans T., Vinje J. Simultaneous recovery of DNA/RNA from diverse microbes. Pathogens. 2015; 4 (2): 335–351. doi: 10.3390/pathogens4020335.Schriewer A., Wehlmann A., Wuertz S. Removing humic acids with DAX‑8 improves qPCR. J Microbiol Methods. 2011; 85 (1): 16–21. doi: 10.1016/j.mimet.2010.12.027.Schriewer A., Wehlmann A., Wuertz S. Removing humic acids with DAX‑8 improves qPCR. J Microbiol Methods. 2011; 85 (1): 16–21. doi: 10.1016/j.mimet.2010.12.027.Hata A., Kitajima M., Katayama H. Resin + gel filtration to mitigate inhibitors in virus quantification. Water Res. 2020; 173: 115536. doi: 10.1016/j.watres.2020.115536.Hata A., Kitajima M., Katayama H. Resin + gel filtration to mitigate inhibitors in virus quantification. Water Res. 2020; 173: 115536. doi: 10.1016/j.watres.2020.115536.Guttman-Bass N., Catalano-Sherman J., Klein B. S. Humic acid interference with virus recovery by membrane filtration. Appl Environ Microbiol. 1986; 51 (5): 883–885.Guttman-Bass N., Catalano-Sherman J., Klein B. S. Humic acid interference with virus recovery by membrane filtration. Appl Environ Microbiol. 1986; 51 (5): 883–885.Bonilla N., Rojas M. I., Cruz G. N. F., Hung S.-H., Rohwer F., Barr J. J. Phage on tap — quick and efficient preparation of high‑titer stocks. PeerJ. 2016; 4: e2261. doi: 10.7717/peerj.2261.Bonilla N., Rojas M. I., Cruz G. N. F., Hung S.-H., Rohwer F., Barr J. J. Phage on tap — quick and efficient preparation of high‑titer stocks. PeerJ. 2016; 4: e2261. doi: 10.7717/peerj.2261.Hatfull G. F. Mycobacteriophages. Microbiol Spectr. 2018; 6 (5). doi: 10.1128/microbiolspec.GPP3-0026-2018.Hatfull G. F. Mycobacteriophages. Microbiol Spectr. 2018; 6 (5). doi: 10.1128/microbiolspec.GPP3-0026-2018.Yang Y., Shen W., Zhong Q., Chen Q. Essential role of calcium in infection by a broad-host-range phage. J Basic Microbiol. 2013; 53 (7): 1–9. doi: 10.1002/jobm.201300051.Yang Y., Shen W., Zhong Q., Chen Q. Essential role of calcium in infection by a broad-host-range phage. J Basic Microbiol. 2013; 53 (7): 1–9. doi: 10.1002/jobm.201300051.Ahmed E. et al. Geochemical constraints on bacteriophage infectivity. bioRxiv. 2023; preprint 2023.04.10.536276.Ahmed E. et al. Geochemical constraints on bacteriophage infectivity. bioRxiv. 2023; preprint 2023.04.10.536276.SEA-PHAGES Program. Phage Discovery Guide. 5th ed. HHMI; 2020. Available at: https://seaphages.org/content/phoenix/.SEA-PHAGES Program. Phage Discovery Guide. 5th ed. HHMI; 2020. Available at: https://seaphages.org/content/phoenix/.Stachurska X., Roszak M., Jabłońska J., Mizielińska M., Nawrotek P. Dounle-layer Agar (DLA) modifications for phage-antibiotic synergy identification. Antibiotics (Basel). 2021; 10 (11): 1306. doi: 10.3390/antibiotics10111306.Stachurska X., Roszak M., Jabłońska J., Mizielińska M., Nawrotek P. Dounle-layer Agar (DLA) modifications for phage-antibiotic synergy identification. Antibiotics (Basel). 2021; 10 (11): 1306. doi: 10.3390/antibiotics10111306.Adams M. H. Bacteriophages. New York: Interscience Publishers; 1959.Adams M. H. Bacteriophages. New York: Interscience Publishers; 1959.Hyman P., Abedon S. T. Practical methods for determining phage growth parameters. Methods Mol Biol. 2009; 501: 175–202. doi: 10.1007/978-1-60327-164-6_18.Hyman P., Abedon S. T. Practical methods for determining phage growth parameters. Methods Mol Biol. 2009; 501: 175–202. doi: 10.1007/978-1-60327-164-6_18.Kutter E., Sulakvelidze A. (eds). Bacteriophages: Biology and Applications. Boca Raton (FL): CRC Press; 2005.Kutter E., Sulakvelidze A. (eds). Bacteriophages: Biology and Applications. Boca Raton (FL): CRC Press; 2005.Pope W. H., Bowman C. A., Russell D. A., Jacobs-Sera D., Asai D. J., Cresawn S. G. et al. Whole genome comparison of a large collection of mycobacteriophages reveals a continuum of phage genetic diversity. eLife. 2015; 4: e06416. doi: 10.7554/eLife.06416.Pope W. H., Bowman C. A., Russell D. A., Jacobs-Sera D., Asai D. J., Cresawn S. G. et al. Whole genome comparison of a large collection of mycobacteriophages reveals a continuum of phage genetic diversity. eLife. 2015; 4: e06416. doi: 10.7554/eLife.06416.Hatfull G. F. Mycobacteriophages: genes and genomes. Annu Rev Microbiol. 2014; 68: 225–243. doi: 10.1146/annurev.micro.112408.134233.Hatfull G. F. Mycobacteriophages: genes and genomes. Annu Rev Microbiol. 2014; 68: 225–243. doi: 10.1146/annurev.micro.112408.134233.Mirzaei M. K., Nilsson A. S. Correction: isolation of phages for phage therapy: a practical approach. Methods Mol Biol. 2015; 1317: 3–14.Mirzaei M. K., Nilsson A. S. Correction: isolation of phages for phage therapy: a practical approach. Methods Mol Biol. 2015; 1317: 3–14.Ackermann H. W. 5500 phages examined in the electron microscope. Arch Virol. 2007; 152 (2): 227–243. doi: 10.1007/s00705-006-0849-1.Ackermann H. W. 5500 phages examined in the electron microscope. Arch Virol. 2007; 152 (2): 227–243. doi: 10.1007/s00705-006-0849-1.Russell D. A. Sequencing, assembling, and finishing complete bacteriophage genomes. In: bacteriophages: Methods and Protocols. Vol 3. Springer; 2018.Russell D. A. Sequencing, assembling, and finishing complete bacteriophage genomes. In: bacteriophages: Methods and Protocols. Vol 3. Springer; 2018.Bankevich A., Nurk S., Antipov D., Gurevich A. A., Dvorkin M., Kulikov A. S. et al. SPAdes genome assembly algorithm. J Comput Biol. 2012; 19 (5): 455–477. doi: 10.1089/cmb.2012.0021.Bankevich A., Nurk S., Antipov D., Gurevich A. A., Dvorkin M., Kulikov A. S. et al. SPAdes genome assembly algorithm. J Comput Biol. 2012; 19 (5): 455–477. doi: 10.1089/cmb.2012.0021.Wick R. R., Judd L. M., Gorrie C. L., Holt K. E. Unicycler resolves assemblies from short/long reads. PLoS Comput Biol. 2017; 13 (6): e1005595. doi: 10.1371/journal.pcbi.1005595.Wick R. R., Judd L. M., Gorrie C. L., Holt K. E. Unicycler resolves assemblies from short/long reads. PLoS Comput Biol. 2017; 13 (6): e1005595. doi: 10.1371/journal.pcbi.1005595.Kolmogorov M., Yuan J., Lin Y., Pevzner P. A. Assembly of long, errorprone reads using repeat graphs. Nat Biotechnol. 2019; 37: 540–546. doi: 10.1038/s41587-019-0072-8.Kolmogorov M., Yuan J., Lin Y., Pevzner P. A. Assembly of long, errorprone reads using repeat graphs. Nat Biotechnol. 2019; 37: 540–546. doi: 10.1038/s41587-019-0072-8.Garneau J. R., Depardieu F., Fortier L. C., Bikard D., Monot M. PhageTerm: a tool for fast and accurate determination of phage termini and packaging mechanism using next-generation sequencing data. Sci Rep. 2017; 7: 8292. doi: 10.1038/s41598-017-07910-5.Garneau J. R., Depardieu F., Fortier L. C., Bikard D., Monot M. PhageTerm: a tool for fast and accurate determination of phage termini and packaging mechanism using next-generation sequencing data. Sci Rep. 2017; 7: 8292. doi: 10.1038/s41598-017-07910-5.McNair K., Zhou C., Dinsdale E. A., Souza B., Edwards R. A. PHANOTATE: gene identification in phage genomes. Bioinformatics. 2019; 35 (22): 4537–4542. doi: 10.1093/bioinformatics/btz265.McNair K., Zhou C., Dinsdale E. A., Souza B., Edwards R. A. PHANOTATE: gene identification in phage genomes. Bioinformatics. 2019; 35 (22): 4537–4542. doi: 10.1093/bioinformatics/btz265.Zimmermann L., Stephens A., Nam S. Z., Rau D., Kubler J., Lozajic M. et al. A completely reimplemented MPI bioinformatics toolkit with a new HHpred server at its core. J Mol Biol. 2018; 430 (15): 2237–2243. doi: 10.1016/j.jmb.2017.12.007.Zimmermann L., Stephens A., Nam S. Z., Rau D., Kubler J., Lozajic M. et al. A completely reimplemented MPI bioinformatics toolkit with a new HHpred server at its core. J Mol Biol. 2018; 430 (15): 2237–2243. doi: 10.1016/j.jmb.2017.12.007.Arndt D., Grant J. R., Marcu A., Sajed T., Pon A., Liang Y., Wishart D. S. PHASTER: improved phage search tool. Nucleic Acids Res. 2016; 44 (W1): W16-W21. doi: 10.1093/nar/gkw387.Arndt D., Grant J. R., Marcu A., Sajed T., Pon A., Liang Y., Wishart D. S. PHASTER: improved phage search tool. Nucleic Acids Res. 2016; 44 (W1): W16-W21. doi: 10.1093/nar/gkw387.Alcock B. P., Raphenya A. R., Lau T. T.Y, Tsang K. K., Bouchard M., Edalatmand A. et al. CARD 2020: antibiotic resistome surveillance. Nucleic Acids Res. 2020; 48 (D1): D517-D525. doi: 10.1093/nar/gkz935.Alcock B. P., Raphenya A. R., Lau T. T.Y, Tsang K. K., Bouchard M., Edalatmand A. et al. CARD 2020: antibiotic resistome surveillance. Nucleic Acids Res. 2020; 48 (D1): D517-D525. doi: 10.1093/nar/gkz935.Bortolaia V., Kaas R. S., Ruppe E., Roberts M. C., Schwarz S., Cattoir V. et al. ResFinder 4.0 for phenotype predictions from genotypes. J Antimicrob Chemother. 2020; 75 (12): 3491–3500. doi: 10.1093/jac/dkaa345.Bortolaia V., Kaas R. S., Ruppe E., Roberts M. C., Schwarz S., Cattoir V. et al. ResFinder 4.0 for phenotype predictions from genotypes. J Antimicrob Chemother. 2020; 75 (12): 3491–3500. doi: 10.1093/jac/dkaa345.Nayfach S., Camargo A. P., Schulz F., Eloe-Fadrosh E., Roux S., Ryrpides N. C. CheckV assesses quality of viral MAGs. Nat Biotechnol. 2021; 39: 578–585. doi: 10.1038/s41587-020-00774-7.Nayfach S., Camargo A. P., Schulz F., Eloe-Fadrosh E., Roux S., Ryrpides N. C. CheckV assesses quality of viral MAGs. Nat Biotechnol. 2021; 39: 578–585. doi: 10.1038/s41587-020-00774-7.Marinelli L. J., Piuri M., Swigonová Z., Balachandran A., Oldfield L. M., van Kessel J. C., Hatfull G. F. BRED: a simple and powerful tool for constructing mutant/recombinant phage genomes. PLoS One. 2008; 3 (12): e3957. doi: 10.1371/journal.pone.0003957.Marinelli L. J., Piuri M., Swigonová Z., Balachandran A., Oldfield L. M., van Kessel J. C., Hatfull G. F. BRED: a simple and powerful tool for constructing mutant/recombinant phage genomes. PLoS One. 2008; 3 (12): e3957. doi: 10.1371/journal.pone.0003957.Pires D. P., Cleto S., Sillankorva S., Azeredo J., Lu T. K. Genetically engineered phages: advances over the last decade. ACS Synth Biol. 2016; 5 (10): 1143–1157. doi: 10.1128/MMBR.00069-15.Pires D. P., Cleto S., Sillankorva S., Azeredo J., Lu T. K. Genetically engineered phages: advances over the last decade. ACS Synth Biol. 2016; 5 (10): 1143–1157. doi: 10.1128/MMBR.00069-15.International Council for Harmonisation (ICH). Q1A (R2) Stability Testing of New Drug Substances and Products. Geneva: ICH; 2003.International Council for Harmonisation (ICH). Q1A (R2) Stability Testing of New Drug Substances and Products. Geneva: ICH; 2003.International Council for Harmonisation (ICH). Q6B Specifications: Test Procedures and Acceptance Criteria for Biotechnological/Biological Products. Geneva: ICH; 1999.International Council for Harmonisation (ICH). Q6B Specifications: Test Procedures and Acceptance Criteria for Biotechnological/Biological Products. Geneva: ICH; 1999.United States Pharmacopeia.<71>Sterility Tests. In: USP-NF. Rockville, MD: United States Pharmacopeial Convention; current edition.United States Pharmacopeia.<71>Sterility Tests. In: USP-NF. Rockville, MD: United States Pharmacopeial Convention; current edition.United States Pharmacopeia.<85>Bacterial Endotoxins Test. In: USP-NF. Rockville, MD: United States Pharmacopeial Convention; current edition.United States Pharmacopeia.<85>Bacterial Endotoxins Test. In: USP-NF. Rockville, MD: United States Pharmacopeial Convention; current edition.Merabishvili M., Pirnay J-P, Verbeken G., Cganishvili N., Tediashvili M., Lashkhi N. et al. Quality‑controlled small-scale production of a well-defined bacteriophage cocktail for use in human clinical trials. PLoS One. 2009; 4 (3): e4944. doi: 10.1371/journal.pone.0004944.Merabishvili M., Pirnay J-P, Verbeken G., Cganishvili N., Tediashvili M., Lashkhi N. et al. Quality‑controlled small-scale production of a well-defined bacteriophage cocktail for use in human clinical trials. PLoS One. 2009; 4 (3): e4944. doi: 10.1371/journal.pone.0004944.International Council for Harmonisation (ICH). Q5C Quality of Biotechnological Products: Stability Testing of Biotechnological/Biological Products. Geneva: ICH; 1995.International Council for Harmonisation (ICH). Q5C Quality of Biotechnological Products: Stability Testing of Biotechnological/Biological Products. Geneva: ICH; 1995.European Commission. EudraLex-Volume 4. EU Guidelines for Good Manufacturing Practice for Medicinal Products for Human and Veterinary Use. Annex 1: Manufacture of Sterile Medicinal Products (rev. 2022). Brussels: European Commission; 2022.European Commission. EudraLex-Volume 4. EU Guidelines for Good Manufacturing Practice for Medicinal Products for Human and Veterinary Use. Annex 1: Manufacture of Sterile Medicinal Products (rev. 2022). Brussels: European Commission; 2022.Federal Agency for Medicines and Health Products (FAMHP, Belgium). Magistral preparation of bacteriophage medicines: framework and quality requirements. Brussels: FAMHP; 2018.Federal Agency for Medicines and Health Products (FAMHP, Belgium). Magistral preparation of bacteriophage medicines: framework and quality requirements. Brussels: FAMHP; 2018.Beinhauerova M., Slana I. Phage amplification assays for detection of Mycobacterium avium subsp. paratuberculosis: a review. Microorganisms. 2021; 9 (2): 237. doi: 10.3390/microorganisms9020237.Beinhauerova M., Slana I. Phage amplification assays for detection of Mycobacterium avium subsp. paratuberculosis: a review. Microorganisms. 2021; 9 (2): 237. doi: 10.3390/microorganisms9020237.Jain P., Hartman T. E., Eisenberg N. et al. Phage Lysin B for detection of Mycobacterium tuberculosis complex: Proof-of-principle. J Clin Microbiol. 2012; 50 (4): 1362–1369. doi: 10.1128/JCM.06138-11.Jain P., Hartman T. E., Eisenberg N. et al. Phage Lysin B for detection of Mycobacterium tuberculosis complex: Proof-of-principle. J Clin Microbiol. 2012; 50 (4): 1362–1369. doi: 10.1128/JCM.06138-11.Piuri M., Jacobs W. R. Jr., Hatfull G. F. Fluoromycobacteriophages for rapid, specific, and sensitive antibiotic susceptibility testing of Mycobacterium tuberculosis. PLoS One. 2009; 4 (3): e4870. doi: 10.1371/journal.pone.0004870.Piuri M., Jacobs W. R. Jr., Hatfull G. F. Fluoromycobacteriophages for rapid, specific, and sensitive antibiotic susceptibility testing of Mycobacterium tuberculosis. PLoS One. 2009; 4 (3): e4870. doi: 10.1371/journal.pone.0004870.Jain P., Hsu T., Arai M. et al. Systems approach identifies gene networks for tuberculosis progression and reveals targets for intervention. J Bacteriol. 2020; 202 (24): e00411-20. doi: 10.1128/JB.00411-20.Jain P., Hsu T., Arai M. et al. Systems approach identifies gene networks for tuberculosis progression and reveals targets for intervention. J Bacteriol. 2020; 202 (24): e00411-20. doi: 10.1128/JB.00411-20.Rajagopalan S., Mehra S., Kumar P. et al. Evaluation of a phage-based test for rapid tuberculosis diagnosis: a multicenter study. J Clin Microbiol. 2025; 63 (9): e00841-25. doi: 10.1128/JCM.00841-25.Rajagopalan S., Mehra S., Kumar P. et al. Evaluation of a phage-based test for rapid tuberculosis diagnosis: a multicenter study. J Clin Microbiol. 2025; 63 (9): e00841-25. doi: 10.1128/JCM.00841-25.Prakash S., Jaiswal N., Jain A. Correlation between phage assay and culture for Mycobacterium tuberculosis detection: A meta-analysis. Int J Tuberc Lung Dis. 2009; 13 (7): 774–779.Prakash S., Jaiswal N., Jain A. Correlation between phage assay and culture for Mycobacterium tuberculosis detection: A meta-analysis. Int J Tuberc Lung Dis. 2009; 13 (7): 774–779.Murthy M. K., Gupta V. K., Maurya A. P., Diagnosis of nontuberculous mycobacterial infections using genomics and artificial intelligence-machine learning approaches: scope, progress and challenges. Front Microbiol. 2025; 16: 1665685. doi: 10.3389/fmicb.2025.1665685.Murthy M. K., Gupta V. K., Maurya A. P., Diagnosis of nontuberculous mycobacterial infections using genomics and artificial intelligence-machine learning approaches: scope, progress and challenges. Front Microbiol. 2025; 16: 1665685. doi: 10.3389/fmicb.2025.1665685.Kortright K. E., Chan B. K., Koff J. L., Turner P. E. Phage therapy: A renewed approach to combat antibiotic‑resistant bacteria. Cell Host Microbe. 2019; 25 (2): 219–232. doi: 10.1016/j.chom.2019.01.014.Kortright K. E., Chan B. K., Koff J. L., Turner P. E. Phage therapy: A renewed approach to combat antibiotic‑resistant bacteria. Cell Host Microbe. 2019; 25 (2): 219–232. doi: 10.1016/j.chom.2019.01.014.Schooley R. T., Biswas B., Gill J. J., Hernandez-Morales A., Lancaster J., Lessor L. et al. Development and use of personalized bacteriophage‑based therapeutic cocktails to treat a patient with a disseminated resistant Acinetobacter baumannii infection. Antimicrob Agents Chemother. 2017; 61 (10): e00954-17. doi: 10.1128/AAC.00954-17.Schooley R. T., Biswas B., Gill J. J., Hernandez-Morales A., Lancaster J., Lessor L. et al. Development and use of personalized bacteriophage‑based therapeutic cocktails to treat a patient with a disseminated resistant Acinetobacter baumannii infection. Antimicrob Agents Chemother. 2017; 61 (10): e00954-17. doi: 10.1128/AAC.00954-17.Chang R. Y. K., Morales S., Chan H. K. Topical liquid formulation of bacteriophages for metered dose delivery to cutaneous wounds. Adv Drug Deliv Rev. 2022; 181: 114080. doi: 10.1016/j.addr.2021.114080.Chang R. Y. K., Morales S., Chan H. K. Topical liquid formulation of bacteriophages for metered dose delivery to cutaneous wounds. Adv Drug Deliv Rev. 2022; 181: 114080. doi: 10.1016/j.addr.2021.114080.Aslam S., Lampley E., Wooten D., Karris M. Y., Benson C., Strathdee S., Schooley R. Lessons learned from the first 10 consecutive cases of intravenous bacteriophage therapy to treat multidrug-resistant bacterial infections at a single center in the US. Open Forum Infect Dis. 2020; 7 (9): ofaa389. doi: 10.1093/ofid/ofaa389.Aslam S., Lampley E., Wooten D., Karris M. Y., Benson C., Strathdee S., Schooley R. Lessons learned from the first 10 consecutive cases of intravenous bacteriophage therapy to treat multidrug-resistant bacterial infections at a single center in the US. Open Forum Infect Dis. 2020; 7 (9): ofaa389. doi: 10.1093/ofid/ofaa389.Abedon S. T. Phage therapy pharmacology: calculating phage dosing. Adv Appl Microbiol. 2011; 77: 1–40. doi: 10.1016/B978-0-12-387044-5.00001-7.Abedon S. T. Phage therapy pharmacology: calculating phage dosing. Adv Appl Microbiol. 2011; 77: 1–40. doi: 10.1016/B978-0-12-387044-5.00001-7.Chan B. K., Sistrom M., Wertz J. E., Kortright K. E., Narayan D., Turner P. E. Phage selection restores antibiotic sensitivity in MDR Pseudomonas aeruginosa. Evol Med Public Health. 2018; 2018 (1): 60–66. doi: 10.1093/emph/eoy005.Chan B. K., Sistrom M., Wertz J. E., Kortright K. E., Narayan D., Turner P. E. Phage selection restores antibiotic sensitivity in MDR Pseudomonas aeruginosa. Evol Med Public Health. 2018; 2018 (1): 60–66. doi: 10.1093/emph/eoy005.Torres-Barceló C., Hochberg M. E. Evolutionary rationale for phages as complements of antibiotics. Trends Microbiol. 2016; 24 (4): 249–256. doi: 10.1016/j.tim.2015.12.011.Torres-Barceló C., Hochberg M. E. Evolutionary rationale for phages as complements of antibiotics. Trends Microbiol. 2016; 24 (4): 249–256. doi: 10.1016/j.tim.2015.12.011.Hodyra-Stefaniak K., Miernikiewicz P., Drapała J., Drab M., Joсczyk-Matysiak E., Lecion D. et al. Mammalian host-versus-phage immune response determines phage fate in vivo. Sci Rep. 2015; 5: 14802. doi: 10.1038/srep14802.Hodyra-Stefaniak K., Miernikiewicz P., Drapała J., Drab M., Joсczyk-Matysiak E., Lecion D. et al. Mammalian host-versus-phage immune response determines phage fate in vivo. Sci Rep. 2015; 5: 14802. doi: 10.1038/srep14802.Grant I. R. Bacteriophage-based methods for the detection of viable Mycobacterium avium subsp. paratuberculosis and their potential for diagnosis of Johne's diseases. Front Vet Sci. 2021; 8: 632498. doi: 10.3389/fvets.2021.632498.Grant I. R. Bacteriophage-based methods for the detection of viable Mycobacterium avium subsp. paratuberculosis and their potential for diagnosis of Johne's diseases. Front Vet Sci. 2021; 8: 632498. doi: 10.3389/fvets.2021.632498.Foddai A., Grant I. R. Bacteriophage‑based methods for detection of viable Mycobacterium avium subsp. paratuberculosis: An update. Appl Microbiol Biotechnol. 2020; 104 (21): 9399—9412. doi: 10.1007/s00253-020-10841-y.Foddai A., Grant I. R. Bacteriophage‑based methods for detection of viable Mycobacterium avium subsp. paratuberculosis: An update. Appl Microbiol Biotechnol. 2020; 104 (21): 9399—9412. doi: 10.1007/s00253-020-10841-y.

The authors declare that there are no conflicts of interest present.