tbjournalТуберкулез и социально значимые заболеванияTuberculosis and socially significant diseases2413-03462413-0354ООО «Ин-Тренд10.54921/2413-0346-2022-10-4-48-57tbjournal-114Research ArticleДИАГНОСТИКА И КЛИНИКА ТУБЕРКУЛЕЗАDIAGNOSIS AND CLINIC OF TUBERCULOSISОценка эффективности молекулярногенетических и бактериологических методов в определении гетерорезистентности Mycobacterium tuberculosis к рифампицинуEvaluation of the effectiveness of molecular genetic and bacteriological methods in determining heteroresistance Mycobacterium tuberculosis to rifampicinИсаковаА. И.IsakovaA. I.

Исакова Александра Ивановна – врач клинической лабораторной диагностики Централизованной бактериологическойлаборатории туберкулеза

107014, г. Москва, ул. Стромынка, д. 10

Тел. +7 (499) 268-70-33, тел./факс +7 (499) 785-20-82

isakovaaleks@gmail.comМихайловаЮ. Д.MikhailovaYu. D.

Михайлова Юлия Дмитриевна – ведущий научный сотрудник отдела проблем лабораторной диагностики туберкулеза, кандидат биологических наук

107014, г. Москва, ул. Стромынка, д. 10

Тел. +7 (499) 268-70-33, тел./факс +7 (499) 785-20-82

juliaisaeva81@rambler.ruГалкинаК. Ю.GalkinaK. Yu.

Галкина Ксения Юрьевна – ведущий научный сотрудник отдела проблем лабораторной диагностики туберкулеза и патоморфологии, кандидат биологических наук

107014, u/ Москва, ул. Стромынка, д. 10

Тел./факс +7 (499) 785-20-82

crazymare@mail.ruНосоваЕ. Ю.NosovaE. Yu.

Носова Елена Юрьевна – ведущий научный сотрудник отдела проблем лабораторной диагностики туберкулеза, доктор медицинских наук

107014, г. Москва, ул. Стромынка, д. 10

Тел. +7 (495) 603-30-33, тел./факс +7 (499) 785-20-82

rna68@rambler.ruСафоноваС. Г.SafonovaS. G.

Сафонова Светлана Григорьевна – заведующая отделом проблем лабораторной диагностики туберкулеза, доктор биологических наук

107014, г. Москва, ул. Стромынка, д. 10

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safonova.s.g@inbox.ruГБУЗ города Москвы «Московский городской научно-практический центр борьбы с туберкулезом Департамента здравоохранения города Москвы»Russian Federation2022301220221043241Copyright © Исакова А.И., Михайлова Ю.Д., Галкина К.Ю., Носова Е.Ю., Сафонова С.Г., 20222022Исакова А.И., Михайлова Ю.Д., Галкина К.Ю., Носова Е.Ю., Сафонова С.Г.Isakova A.I., Mikhailova Y.D., Galkina K.Y., Nosova E.Y., Safonova S.G.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.tb-journal.ru/jour/article/view/114

Для оценки эффективности молекулярно-генетических тестов и бактериологических методов по определению гетерорезистентной к рифампицину (R) популяции M. tuberculosis (МБТ) проанализированы бактериальные суспензии (смеси), содержащие различные пропорции устойчивых и чувствительных штаммов.

Установлено, что эффективность определения устойчивой к R популяции МБТ в смесях зависит от используемого в тестсистеме способа детекции мутаций. Тест-системы Genotype MTBDRplus, «ТБ-ТЕСТ» и «ТБ-БИОЧИП-1» показали одинаковые результаты выявления устойчивой популяции с мутацией Ser531Leu. Genotype MTBDRplus эффективнее определяет замены His526Tyr, His526Asp, Asp516Val и менее эффективно, по сравнению с «ТБ-ТЕСТ» и «ТБ-БИОЧИП-1» – мутации, определяемые по отсутствию зондов «дикого» типа. Эффективность бактериологического обнаружения резистентной популяции зависит от сроков культивирования. Метод серийных микроразведений в жидкой среде Миддлбрука 7H9 наиболее эффективный при выявлении устойчивости к Rif в гетерорезистентной популяции МБТ при культивировании посевов 21 день по сравнению с автоматизированной системой BACTEC™ MGIT™ 960 и методом абсолютных концентраций на плотной среде Л-Й при культивировании посевов 28 дней. Отмечено, что доля выявляемых устойчивых штаммов МБТ с помощью Bactec MGIT 960, методом абсолютных концентраций на плотной среде Л-Й при культивировании посевов 28 дней и методом серийных микроразведений при культивировании посевов 14 дней существенно не различалась и составила 45,1, 44,1 и 38,9 соответственно.

We analyzed bacterial suspensions (mixtures) containing various proportions of resistant and susceptible strains to assess the effectiveness of molecular genetic tests and bacteriological methods for determination of the heteroresistant population of M. tuberculosis (MTB) to rifampicin (RIF). We established that the efficiency of determining the RIF-resistant MTB population in mixtures depends on the method of mutation detection used in the test system. The Genotype MTBDRplus, TB-TEST, and TB-BIOCHIP-1 tests showed the same results in detecting a resistant population with the Ser531Leu mutation. Genotype MTBDRplus more effectively detected His526Tyr, His526Asp, Asp516Val substitutions and less efficiently detected mutations determined by the absence of wild-type probes compared to TB-TEST and TB-BIOCHIP-1 tests.

The efficiency of bacteriological detection of the resistant MTB population depends on the terms of cultivation. The method of serial microdilutions in the Middlebrook 7H9 liquid medium is the most effective in detecting RIF-resistance in the heteroresistant MTB population with the MTB cultivation for 21 days in comparison with the automated system Bactec MGIT 960 and the method of absolute concentrations on the L-J solid medium with the MTB cultivation for 28 days. It was noted that the proportion of resistant MTB strains detectection by Bactec MGIT 960, the method of absolute concentrations on a solid L-J medium for 28 days and by the method of serial microdilutions for 14 days did not differ significantly and amounted to 45.1; 44.1 and 38.9 respectively

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