pmjТихоокеанский медицинский журналPacific Medical Journal1609-1175TGMUpmj-606Research ArticleОРИГИНАЛЬНЫЕ ИССЛЕДОВАНИЯORIGINAL RESEARCHESВыявление мутаций устойчивости к макролидам в гене 23S рРНК Mycoplasma pneumoniae с помощью полимеразной цепной реакции в режиме реального времениDetection of macrolide-resistance mutations in 23S rRNA gene of Mycoplasma pneumoniae using a novel real-time PCR assayЭйдельштейнИ. А.EdelsteinI. A.inna.edelstein@antibiotic.ruЭйдельштейнМ. В.EdelsteinM. V.noemail@neicon.ruРомановА. В.RomanovA. V.noemail@neicon.ruРачинаС. А.RatchinaS. A.noemail@neicon.ruЯцышинаС. Б.YatsyshinaS. B.noemail@neicon.ruРаковскаяИ. В.RakovskajaI. V.noemail@neicon.ruКозловР. С.KozlovR. S.noemail@neicon.ruНИИ антимикробной химиотерапии Смоленской государственной медицинской академииРоссияInstitute of Antimicrobial Chemotherapy Smolensk State Medical AcademyRussian FederationЦентральный научно-исследовательский институт эпидемиологииРоссияCentral Research Institute for EpidemiologyRussian FederationНИИ эпидемиологии и микробиологии имени Н.Ф. Гамалеи РАМНРоссияResearch Institute of Epidemiology and Microbiology named under N.F. GamaleyaRussian Federation201528032015016366Copyright © Эйдельштейн И.А., Эйдельштейн М.В., Романов А.В., Рачина С.А., Яцышина С.Б., Раковская И.В., Козлов Р.С., 20152015Эйдельштейн И.А., Эйдельштейн М.В., Романов А.В., Рачина С.А., Яцышина С.Б., Раковская И.В., Козлов Р.С.Edelstein I.A., Edelstein M.V., Romanov A.V., Ratchina S.A., Yatsyshina S.B., Rakovskaja I.V., Kozlov R.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.tmj-vgmu.ru/jour/article/view/606

В последнее время данные зарубежных публикаций свидетельствуют о появлении и распространении устойчивости Mycoplasma pneumoniae к макролидным антибиотикам, которая связана с мутациями в гене 23S рРНК, главным образом, в позициях 2063, 2064 и 2617. Выявление соответствующих однонуклеотидных замен позволяет эффективно предсказать фенотип устойчивости к макролидам, однако, используемые с этой целью методы являются трудоемкими и дорогостоящими. Исследование посвящено разработке и валидации нового метода определения мутаций, ассоциированных с устойчивостью к макролидам у M. pneumoniae, а также его использованию для анализа клинических образцов. Показана возможность выявления и дифференциации различных мутаций в позициях 2063, 2064 и 2617 гена 23S рРНК в формате однопробирочной мультиплексной полимеразной цепной реакции в режиме реального времени с последующим анализом кривых плавления флуоресцентно-меченых зондов. При исследовании клинических образцов во всех случаях обнаружены последовательности 23S рДНК M. pneumoniae «дикого типа», характерные для фенотипа чувствительности к макролидам.

Background. Mycoplasma pneumoniae is a widespread pathogen of the respiratory tract diseases. In recent years, the data of foreign publications indicate the appearance and spread of resistant M. pneumoniae to macrolide antibiotics, which is associated with mutations in the 23S pRNA gene, mainly in positions 2063, 2064 and 2617. Methods. Studied 146 clinical samples obtained in 2006-2013. of patients with infections of the lower respiratory tract. Primary screening was done for the presence of DNA material M. pneumonia based on polymerase chain reaction (PCR) in real time. Results. The possibility of detecting and differentiation of various mutations in positions 2063, 2064 and 2617 in the 23S pRNA gene format single sample multiplex PCR in real time with subsequent melting curve analysis of fluorescently-labeled probes. In the study of clinical samples in all cases been detected 23S pDNA sequences of M. pneumoniae “wild-type” typical for phenotype of sensitivity to macrolides. Conclusions. Detection of single nucleotide corresponding replacements allows effective predict macrolide resistance phenotype, but used for this purpose methods are laborious and expensive. At present data on the prevalence and mechanisms of macrolide resistance in strains of M. pneumoniae in Russia are absent. The present study focuses on the development and validation of a new method for the determination of mutations associated with resistance to mac-rolides in M. pneumoniae, and also its use for the analysis of clinical samples in patients with lower respiratory tract infections.

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