antibioticsАнтибиотики и ХимиотерапияAntibiot Khimioter = Antibiotics and Chemotherapy0235-2990ООО «Издательство ОКИ»10.37489/0235-2990-2024-69-9-10-17-23KUQACMantibiotics-1181Research ArticleЭКСПЕРИМЕНТАЛЬНЫЕ ИССЛЕДОВАНИЯEXPERIMENTAL STUDIESПрименение секвенирования по Сэнгеру в этиологической диагностике бактериальных осложнений в стационареUse of Sanger Sequencing in Etiological Diagnostics of Bacterial Complications in Hospital Environmenthttps://orcid.org/0000-0002-1000-2240БаранцевичН. Е.BarantsevichN. E.

Баранцевич Наталья Евгеньевна — научный сотрудник

AuthorID: 785835. Scopus Author ID: 55880381000

Санкт-Петербург

Natalia E. Barantsevich — Research fellow

AuthorID: 785835. Scopus Author ID: 55880381000

Saint-Petersburg

https://orcid.org/0000-0002-4800-3345БаранцевичЕ. П.BarantsevichE. P.

Баранцевич Елена Петровна — д. м. н., Заведующая научно-исследовательским отделом микробиологии и внутрибольничных инфекций

AuthorID: 268934. Scopus Author ID: 6601955793. WOS Research ID S-1455-2016

Санкт-Петербург

Elena P. Barantsevich — D. Sc. in Medicine, Head of the Research Department of Microbiology and Nosocomial Infections

AuthorID: 268934. Scopus Author ID: 6601955793. WOS Research ID S-1455-2016

Saint-Petersburg

lenabara2003@mail.ruФедеральное государственное бюджетное учреждение «Национальный медицинский исследовательский центр им. В. А. Алмазова» Министерства здравоохранения Российской ФедерацииРоссияAlmazov National Medical Research CentreRussian Federation202425102024699-101723Copyright © Баранцевич Н.Е., Баранцевич Е.П., 20252025Баранцевич Н.Е., Баранцевич Е.П.Barantsevich N.E., Barantsevich E.P.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/1181

Актуальность. Необходимость точной видовой идентификации микроорганизмов, вызывающих инфекционные осложнения у госпитализированных пациентов, в современном здравоохранении не вызывает сомнений. Цель. Определение возможности применения секвенирования по Сенгеру при рутинном микробиологическом обследовании пациентов в клинике внутренних болезней для повышения качества этиологической диагностики бактериальных осложнений. Материал и методы. Исследовали клинические изоляты микроорганизмов, выделенных у пациентов многопрофильного медицинского центра. Применяли классические микробиологические методы посева и идентификации культур, секвенирование по Сэнгеру. Результаты. Идентификация по Сэнгеру с применением системы MicroSeq (Applied Biosystems, США) обеспечила идентификацию всех 231 исследованных изолятов бактерий — возбудителей нозокомиальных инфекций. Для дифференциальной диагностики стрептококков и коагулазонегативных стафилококков в некоторых случаях, когда известные последовательности первых 500 нуклеотидов гена 16S рРНК двух видов различались на 1–2 нуклеотида, повышение дискриминационного уровня видовой идентификации до 100% позволяло провести валидное определение видовой принадлежности изучаемого микроорганизма. Фенотипические методы не позволили выявить значительную часть видов (25,9%) возбудителей нозокомиальных инфекций и лишь 8 (13,8%) из них идентифицировали достоверно во всех случаях. Применение секвенирования по Сэнгеру для идентификации бактерий привело к долговременному эффекту, связанному с повышением квалификации врачей-лаборантов, улучшением дискриминационных возможностей визуальной оценки макроморфологии бактериальных культур, что важно для выявления всех видов микроорганизмов, присутствующих в биосубстратах. Заключение. Метод секвенирования по Сэнгеру является высокоэффективным и достаточно экономичным, по сравнению с широко применяемыми в клинической практике панелями биохимических тестов, методом «золотого стандарта» в этиологической диагностике бактериальных осложнений в клинике внутренних болезней.

Background. The precise identification of microorganism species that cause infectious complications in hospitalized patients is beyond doubt relevant in modern healthcare. The aim of this study was to determine the possibility of using Sanger sequencing in routine microbiological examination of patients in an Internal Medicine Clinic to improve the quality of etiological diagnosis of bacterial complications. Material and methods. Clinical isolates of microorganisms isolated from patients of a multidisciplinary medical center were studied. The study used classical microbiological methods of seeding and identification of cultures, as well as Sanger sequencing. Results. Sanger identification using the MicroSeq system (Applied Biosystems, USA) ensured identification of all 231 studied bacterial isolates – causative agents of nosocomial infections. For differential diagnostics of streptococci and coagulase-negative staphylococci, in some cases, when the known sequences of the first 500 nucleotides of the 16S rRNA gene of two species differed by 1–2 nucleotides, increasing the discrimination level of species identification to 100% allowed valid determination of the species affiliation of the studied microorganism. Phenotypic methods failed to identify a significant proportion of species (25.9%) of nosocomial infection pathogens, and only 8 (13.8%) of them were reliably identified in all cases. The use of Sanger sequencing to identify bacteria led to a long-term effect associated with improved qualifications of laboratory doctors, and enhanced discriminatory capabilities of visual assessment of the macromorphology of bacterial cultures, which is important for identifying all types of microorganisms present in biosubstrates. Conclusion. The Sanger sequencing method is highly efficient and quite cost effective, compared to the biochemical test panels widely used in clinical practice — the «gold standard» method in the etiological diagnosis of bacterial complications in the clinic of internal diseases.

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