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The use of the lytic D29 mycobacteriophages for accelerated determination of the drug sensitivity of tuberculosis mycobacteria to first and second-line anti-TB drugs in clinical isolates obtained after initial cultivation

Mikhail Vladimirsky, Marina Lapenkova, Yulia Alyapkina, Irina Vasilyeva, Greta Arustamova, Pavel Philippov
European Respiratory Journal 2020 56: 5296; DOI: 10.1183/13993003.congress-2020.5296
Mikhail Vladimirsky
1National Medical Research Center for Phthisiopulmonology and Infection Diseases of the Ministry Health of Russia, Moscow, Russian Federation
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  • For correspondence: mvladimirskij@mail.ru
Marina Lapenkova
1National Medical Research Center for Phthisiopulmonology and Infection Diseases of the Ministry Health of Russia, Moscow, Russian Federation
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Yulia Alyapkina
1National Medical Research Center for Phthisiopulmonology and Infection Diseases of the Ministry Health of Russia, Moscow, Russian Federation
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Irina Vasilyeva
1National Medical Research Center for Phthisiopulmonology and Infection Diseases of the Ministry Health of Russia, Moscow, Russian Federation
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Greta Arustamova
1National Medical Research Center for Phthisiopulmonology and Infection Diseases of the Ministry Health of Russia, Moscow, Russian Federation
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Pavel Philippov
2Moscow regional TB dispensary, Moscow, Russian Federation
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Abstract

Introduction: Accelerating and improvement of determining the drug sensitivity of tuberculosis mycobacteria (MTB) in clinical isolates is an urgent problem in the treatment of tuberculosis.

Aims and Objectives: To develop a new culture format for the study of drug sensitivity of MTB using lytic mycobacteriophage and compare the results with studies in the MGIT Bactec system.

Methods: Middlebrook 7H9 liquid nutrient medium with 1mM CaCL2 and OADC enrichment in a 24-well plate format is used. 200 μl of MTB culture, obtained after initial cultivation in the MGIT Bactec system and 800 μl of the culture medium is added to the plate wells. Antibiotics at critical concentrations were added to the plate wells with the exception of control wells (without antibiotics) and incubated for 48 hours. 50 μl of a lytic mycobacteriophage D 29 (107 pfu/ml) was added to each well and incubated for another 48 hours. DNA was isolated and analyze MTB single-copy RegX gene and a specific DNA fragment of D29 using quantitative real-time PCR. The results of the determination of drug sensitivity were positive with differences in the threshold cycle of test samples and a control sample of at least three cycles.

Results: When comparing the used method and the MGIT Bactec system of 108 clinical isolates, the coincidence of the results of determining drug sensitivity was shown in 98% and 97.5% of cases, respectively.

Conclusions: The method for determining drug sensitivity using lytic mycobacteriophage was accelerated up to 5 days, simple to implement and cost-effective.

  • MDR-TB (multidrug-resistant tuberculosis)
  • Experimental approaches
  • Bacteria

Footnotes

Cite this article as: European Respiratory Journal 2020; 56: Suppl. 64, 5296.

This abstract was presented at the 2020 ERS International Congress, in session “Respiratory viruses in the "pre COVID-19" era”.

This is an ERS International Congress abstract. No full-text version is available. Further material to accompany this abstract may be available at www.ers-education.org (ERS member access only).

  • Copyright ©the authors 2020
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The use of the lytic D29 mycobacteriophages for accelerated determination of the drug sensitivity of tuberculosis mycobacteria to first and second-line anti-TB drugs in clinical isolates obtained after initial cultivation
Mikhail Vladimirsky, Marina Lapenkova, Yulia Alyapkina, Irina Vasilyeva, Greta Arustamova, Pavel Philippov
European Respiratory Journal Sep 2020, 56 (suppl 64) 5296; DOI: 10.1183/13993003.congress-2020.5296

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The use of the lytic D29 mycobacteriophages for accelerated determination of the drug sensitivity of tuberculosis mycobacteria to first and second-line anti-TB drugs in clinical isolates obtained after initial cultivation
Mikhail Vladimirsky, Marina Lapenkova, Yulia Alyapkina, Irina Vasilyeva, Greta Arustamova, Pavel Philippov
European Respiratory Journal Sep 2020, 56 (suppl 64) 5296; DOI: 10.1183/13993003.congress-2020.5296
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