mesЭкстремальная биомедицинаExtreme Medicine3033-89643033-8972Centre for Strategic Planning of the Federal Medical and Biological Agency10.47183/mes.2021.035mes-153Research ArticleОРИГИНАЛЬНОЕ ИССЛЕДОВАНИЕORIGINAL RESEARCHМолекулярно-генетическая характеристика трех новых бактериофагов Klebsiella pneumoniae, перспективных для применения в фаговой терапииMolecular genetic characterization of three new Klebsiella pneumoniae bacteriophages suitable for phage therapyГородничевР. Б.GorodnichevR. B.

Роман Борисович Городничев

ул. Малая Пироговская, д. 1а, г. Москва, 119435

Roman B. Gorodnichev

Malaya Pirogovskaya, 1а, Moscow, 119435

gorodnichev.r.b@gmail.comКорниенкоМ. А.KornienkoM. A.

ул. Малая Пироговская, д. 1а, г. Москва, 119435

Malaya Pirogovskaya, 1а, Moscow, 119435

КупцовН. С.KuptsovN. S.

ул. Малая Пироговская, д. 1а, г. Москва, 119435

Malaya Pirogovskaya, 1а, Moscow, 119435

МалаховаМ. В.MalakhovaM. V.

ул. Малая Пироговская, д. 1а, г. Москва, 119435

Malaya Pirogovskaya, 1а, Moscow, 119435

БеспятыхД. А.BespiatykhD. A.

ул. Малая Пироговская, д. 1а, г. Москва, 119435

Malaya Pirogovskaya, 1а, Moscow, 119435

ВеселовскийВ. А.VeselovskyV. A.

ул. Малая Пироговская, д. 1а, г. Москва, 119435

Malaya Pirogovskaya, 1а, Moscow, 119435

ШитиковЕ. А.ShitikovE. A.

ул. Малая Пироговская, д. 1а, г. Москва, 119435

Malaya Pirogovskaya, 1а, Moscow, 119435

ИльинаЕ. Н.IlinaE. N.

ул. Малая Пироговская, д. 1а, г. Москва, 119435

Malaya Pirogovskaya, 1а, Moscow, 119435

Федеральный научно-клинический центр физико-химической медицины Федерального медико-биологического агентстваРоссияFederal Research and Clinical Center of Physical-Chemical Medicine of Federal Medical Biological AgencyRussian Federation2021271020242339097Copyright © Городничев Р.Б., Корниенко М.А., Купцов Н.С., Малахова М.В., Беспятых Д.А., Веселовский В.А., Шитиков Е.А., Ильина Е.Н., 20242024Городничев Р.Б., Корниенко М.А., Купцов Н.С., Малахова М.В., Беспятых Д.А., Веселовский В.А., Шитиков Е.А., Ильина Е.Н.Gorodnichev R.B., Kornienko M.A., Kuptsov N.S., Malakhova M.V., Bespiatykh D.A., Veselovsky V.A., Shitikov E.A., Ilina E.N.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.extrememedicine.ru/jour/article/view/153

Бактерия Klebsiella pneumoniae способна вызывать широкий спектр внутрибольничных инфекций человека, ассоциированных с антибиотикорезистентностью и высокой смертностью. Одна из перспективных альтернатив применению антибиотиков для лечения таких инфекций — терапия вирулентными бактериофагами. Целью работы было выделить из внешней среды вирулентные бактериофаги, эффективные против актуальных клинических штаммов K. pneumoniae, и дать их молекулярно-генетическую характеристику. Бактериофаги выделяли из проб речной воды методом накопительных культур. Полногеномное секвенирование бактериофагов выполняли на платформе MiSeq (Illumina). Выделено и описано три новых бактериофага K. pneumoniae, принадлежащих к семействам Autographiviridae (vB_KpnP_NER40, GenBank MZ602146) и Myoviridae (vB_KpnM_VIK251, GenBank MZ602147; vB_KpnM_FRZ284, GenBank MZ602148). На коллекции из 105 клинических штаммов K. pneumoniae установлено, что бактериофаги vB_KpnP_NER40 и vB_KpnM_VIK251 обладают узким спектром литической активности (22% и 11%), ограниченным штаммами с капсульным типом К2 и К20 соответственно. Бактериофаг vB_KpnM_FRZ284, напротив, имел широкий спектр литической активности (37%), вызывая лизис штаммов с различным типом капсульного полисахарида. Фаги обладают строго вирулентной природой и не несут в составе генома гены интеграз, токсинов или факторов патогенности. В составе геномов капсулоспецифичных бактериофагов vB_KpnP_NER40 и vB_KpnM_VIK251 обнаружены гены деполимераз, кодирующие потенциальные рецепторсвязывающие белки. Коктейль из трех бактериофагов лизировал около 65% штаммов исследуемой коллекции K. рneumoniae и потенциально применим в терапевтических целях.

The Klebsiella pneumoniae bacterium is capable of causing the broad range of human nosocomial infections associated with antibiotic resistance and high mortality. Virulent bacteriophage therapy is one of the promising alternatives to antibiotic treatment of such infections. The study was aimed to isolate virulent bacteriophages effective against the relevant clinical K. pneumoniae strains, and to perform the molecular genetic characterization of these phages. Bacteriophages were isolated from the river water samples using the enrichment method. The whole-genome sequencing was performed on the MiSeq platform (Illumina). Three novel K. pneumoniae bacteriophages belonging to families Autographiviridae (vB_KpnP_NER40, GenBank MZ602146) and Myoviridae (vB_KpnM_VIK251, GenBank MZ602147; vB_KpnM_FRZ284, GenBank MZ602148) have been isolated and characterized. On the collection of 105 K. pneumoniae clinical strains, it has been found that bacteriophages vB_KpnP_NER40 and vB_KpnM_VIK251 have a narrow lytic spectrum (22% and 11%), which is limited to strains of the capsular types К2 and К20 respectively. In contrast, bacteriophage vB_KpnM_FRZ284 has a broad lytic spectrum (37%), causing the lysis of strains with different types of capsular polysaccharide. The phages are strictly virulent and have no genes encoding integrases, toxins or pathogenicity factors in their genomes. Genes of depolymerases, encoding the potential receptor binding proteins, have been found in the genomes of the capsular-specific bacteriophages vB_KpnP_NER40 and vB_KpnM_VIK251. The cocktail of three bacteriophages has lysed about 65% of the studied collection of K. рneumoniae strain and is potentially applicable for therapeutic purposes.

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