Аннотация
Актуальность. Изучение антибиотикоустойчивости микроорганизмов представляет большой интерес для науки и клинической практики. Приобретение устойчивости к антимикробным препаратам (АМП) микроорганизмов является одним из примеров эволюции. Необходимо изучение общих принципов, механизмов, связанных с появлением устойчивости к АМП A. baumannii.
Цель — оценить чувствительность к АМП и определить наличие генов резистентности к β-лактамным АМП A. baumannii, выделенных в г. Ростов-на-Дону c 2020 по 2022 год.
Материал и методы. Штаммы получены от пациентов с внебольничными и внутрибольничными пневмониями, из смывов с объектов окружающей среды, больниц г. Ростов-на-Дону, в период с 2020 по 2022 год. Культивирование микроорганизмов проводили бактериологическим методом. Видовая принадлежность подтверждена биохимическими методами и MALDI-ToF-масс-спектрометрией. Изучение устойчивости штаммов к АМП проводили диско-диффузионным методом. Поиск генов устойчивости к β-лактамным АМП определяли методом биоинформационного анализа полногеномных сиквенсов программой ResystanceAnalyzer, и методом ПЦР.
Результаты. При исследовании проб, полученных от пациентов, выделены представители рода Acinetobacter, преобладал вид A. baumannii — 72%. Среди образцов смывов с объектов окружающей среды A. baumannii выявлен в 16,2% случаев. Большинство штаммов, выделенных из образцов мокроты, обладали устойчивостью к группе β-лактамных АМП. Методом ПЦР у 9 штаммов выявлены гены группы ОХА-48. Анализ нуклеотидных последовательностей выделенных штаммов A. baumannii, показал наличие генов, кодирующих 7 типов β-лактамаз.
Обсуждение. Штаммы A. baumannii показали полную либо частичную резистентность к АМП, что указывает на наличие большого количества различных механизмов, которые лежат в основе устойчивости к антибиотикам. Выявлены гены резистентности к β-лактамным антибиотикам, благодаря использованию метода ПЦР и полногеномного секвенирования. Было обнаружено, что гены устойчивости к β-лактамам присутствуют в различных сочетаниях генов: bla Adc; bla Oxa; bla AmpC; bla Ctx-M; bla Carb; bla Tem; bla Ndm. По сравнению с ПЦР полногеномное секвенирование помогает обнаружить более широкий диапазон детерминант антибиотикоустойчивости.
Annotation
Introduction. The study of antibiotic resistance of microorganisms is of great interest for science and clinical practice. The acquisition of resistance to antibacterial drugs of microorganisms is one of the examples of evolution. It is necessary to study the general principles and mechanisms associated with the emergence of antibiotic resistance to A. baumannii.
Objective. To evaluate the sensitivity to antibacterial drugs and determine the presence of A. baumannii beta-lactam antibiotic resistance genes isolated in Rostov-on-Don from 2020 to 2022.
Material and methods. The strains were obtained from patients with community-acquired and nosocomial pneumonia, as well as from flushes from environmental facilities, hospitals in Rostov-on-Don, in the period from 2020 to 2022. The cultivation of microorganisms was carried out by the bacteriological method. The species identity was confirmed by biochemical methods and MALDI-ToF mass spectrometry. The study of the resistance of strains to antibacterial drugs was carried out by the disco diffusion method. The search for beta-lactam antibiotic resistance genes was determined by bioinformatic analysis of genome-wide sequences using the ResystanceAnalyzer program, as well as by PCR.
Results. In the study of samples obtained from patients, representatives of the genus Acinetobacter were isolated, mainly the species A. baumannii was isolated – 72%. Among the samples of flushes from environmental objects, A. baumannii was detected in 16.2% of cases. Most of the strain isolated from sputum samples were resistant to the group of beta-lactam antibiotics. The PCR method in 9 strains revealed the genes of the OXA-48 group. The analysis of nucleotide sequences isolated from A. baumannii strains showed the presence of genes encoding 7 types of β-lactamases.
Discussion. A. baumannii strains have shown full or partial resistance to antimicrobial drugs, which indicates the presence of a large number of different mechanisms that underlie antibiotic resistance. Beta-lactam antibiotic resistance genes have been identified through the use of PCR and genome-wide sequencing. It was found that β-lactam resistance genes are present in various gene combinations: bla Adc; bla Oxa; bla AmpC; bla Ctx-M; bla Carb; bla Tem; bla Ndm. Compared with PCR, genome-wide sequencing helps to detect a wider range of determinants of antibiotic resistance.
Conclusion. A. baumannii shows high resistance to antibiotics OXA beta-lactamase genes were most often present.
Key words: Acinetobacter baumannii; pneumonia; antibiotic resistance genes; antibacterial drugs
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