tibljТуберкулез и болезни легкихTuberculosis and Lung Diseases2075-12302542-1506Медицинские знания и технологии10.58838/2075-1230-2023-101-3-69-77tiblj-1733Research ArticleОРИГИНАЛЬНЫЕ СТАТЬИORIGINAL ARTICLESГенетические детерминанты устойчивости МБТ к рифампицину, не включенные в состав отечественных молекулярно-генетических тест-системGenetic Determinants of Rifampicin Resistance of Mycobacterium tuberculosis not Included in the Russian Molecular Genetic Test SystemsМазуринаЕ. А.MazurinaE. A.Мазурина Елена Александровна, Младший научный сотрудник научно-исследовательского отдела микробиологии и доклинических исследований

620039, Свердловская область, г. Екатеринбург, ул. 22-го Партсъезда, д. 50 

Elena A. Mazurina, Junior Researcher of Research Department  of Microbiology and Preclinical Studies 

50, XXII Parts'ezda St., Yekaterinburg, Sverdlovsk Region, 620039 

l.mazurina2011@ya.ruУмпелеваТ. В.UmpelevaT. V.Умпелева Татьяна Валерьевна, Ведущий научный сотрудник научно-исследовательского отдела микробиологии и доклинических исследований  

620039, Свердловская область, г. Екатеринбург, ул. 22-го Партсъезда, д. 50 

Tatiana V. Umpeleva, Leading Researcher of Research Department  of Microbiology and Preclinical Studies 

50, XXII Parts'ezda St., Yekaterinburg, Sverdlovsk Region, 620039 

tumpeleva@ya.ruГолубеваЛ. А.GolubevaL. A.

620039, Свердловская область, г. Екатеринбург, ул. 22-го Партсъезда, д. 50 

50, XXII Parts'ezda St., Yekaterinburg, Sverdlovsk Region, 620039 

ЛавренчукЛ. С.LavrenchukL. S.Лавренчук Леонид Сергеевич, Младший научный сотрудник научно-исследовательского отдела микробиологии и доклинических исследований 

620039, Свердловская область, г. Екатеринбург, ул. 22-го Партсъезда, д. 50 

Leonid S. Lavrenchuk, Junior Researcher of Research Department  of Microbiology and Preclinical Studies

50, XXII Parts'ezda St., Yekaterinburg, Sverdlovsk Region, 620039 

ВахрушеваД. В.VakhrushevaD. V.Вахрушева Диана Владимировна, Заведующая научно-исследовательским отделом микробиологии и доклинических исследований  

620039, Свердловская область, г. Екатеринбург, ул. 22-го Партсъезда, д. 50 

Diana V. Vakhrusheva, Head of Research Department of Microbiology and Preclinical Studies

50, XXII Parts'ezda St., Yekaterinburg, Sverdlovsk Region, 620039 

vakhrusheva@urniif.ruВасильеваИ. А.VasilyevaI. A.Васильева Ирина Анатольевна, Д. м. н., профессор, директор, заведующая кафедрой фтизиатрии  

127473, Москва, ул. Достоевского, д. 4 

Irina A. Vasilyeva, Doctor of Medical Sciences, Professor, Director, Head of Phthisiology Department

4, Dostoevsky St., Moscow, 127473 

Уральский научно-исследовательский институт фтизиопульмонологии – филиал ФГБУ «НМИЦ ФПИ» МЗ РФРоссияUral Phthisiopulmonology Research Institute – a Branch of National Medical Research Center of Phthisiopulmonology and Infectious Diseases, Russian Ministry of HealthRussian FederationФГБУ «Национальный медицинский исследовательский центр фтизиопульмонологии и инфекционных заболеваний» МЗ РФРоссияNational Medical Research Center of Phthisiopulmonology and Infectious Diseases, Russian Ministry of HealthRussian Federation202304072023101369–77Copyright © Мазурина Е.А., Умпелева Т.В., Голубева Л.А., Лавренчук Л.С., Вахрушева Д.В., Васильева И.А., 20232023Мазурина Е.А., Умпелева Т.В., Голубева Л.А., Лавренчук Л.С., Вахрушева Д.В., Васильева И.А.Mazurina E.A., Umpeleva T.V., Golubeva L.A., Lavrenchuk L.S., Vakhrusheva D.V., Vasilyeva I.A.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.tibl-journal.com/jour/article/view/1733Цель исследования

Цель исследования: изучить генетические детерминанты лекарственной устойчивости МБТ к рифампицину путем секвенирования RRDR гена rpoB.

Материалы и методы

Материалы и методы. В исследование включен 651 изолят M. tuberculosis, тесты на лекарственную устойчивость (ТЛЧ) которых были проведены генотипическими методами (гТЛЧ): TB-TEST (БИОЧИП-ИМБ, Россия) и «Амплитуб-МЛУ-РВ»

(НПК Синтол, Россия) и фенотипическими методами (фТЛЧ): метод абсолютных концентраций, система BACTEC MGIT 960, набор Sensititre Myco TB. Для 20 изолятов с расхождениями результатов фТЛЧ и гТЛЧ проведено секвенирование методом Сэнгера RRDR гена rpoB.

Результаты

Результаты. В результате секвенирования обнаружено два варианта мутаций в регионе RRDR гена rpoB, которые не детектируются тест-системами ТБ-ТЕСТ и Амблитуб МЛУ-РВ, но достоверно ассоциированы с устойчивостью МБТ к рифампицину. Наиболее распространенным вариантом явилась вставка трех нуклеотидов (TTC), кодирующих аминокислоту фенилаланин в позиции 434 кодона (1296-1300insTTC).

The objective

The objective: to study the genetic determinants of rifampicin resistance of Mycobacterium tuberculosis by sequencing the RRDR rpoB gene.

Subjects and Methods

Subjects and Methods. The study included 651 M. tuberculosis isolates that were tested for drug suceptibility (DST) by genotypic methods (gDST) TB-TEST (BIOCHIP-IMB, Russia), and Amplitube-MDR-RV (NPK Sintol, Russia) and phenotypic methods (phDST): the absolute concentration method, BACTEC MGIT 960 system, and Sensititre Myco TB kit. Sanger RRDR sequencing of the rpoB gene was performed for 20 isolates with discrepancies in the results of phDST and gDST.

Results

Results. As a result of sequencing, two variants of mutations in the RRDR region of the rpoB gene were found, which were not detected by TB-TEST and Amblitub MDR-RV test systems, however were strongly associated with rifampicin resistance of Mycobacterium tuberculosis. The most common variant was the insertion of three nucleotides (TTC) encoding the amino acid phenylalanine at position 434 of the codon (1296-1300insTTC).

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