Аннотация
Диагностика этиологических агентов хронического пародонтита грамотрицательных анаэробных бактерий Porphyromonas
gingivalis затруднена из-за высокой требовательности данного вида в дополнительных факторах роста.
Цель исследования: повышение чувствительности питательной среды и сокращение срока выделения чистой культуры P.
gingivalis. В работе использованы культуральный и молекулярно-генетический методы. Подобрана питательная среда Бруцеллагар с учётом культуральных особенностей данного пародонтопатогена с добавлением чёрного альбумина и менахинона
(витамин K2
). На разработанной питательной среде получен рост мелких округлых слизистых тёмнопигментированных
колоний, идентифицированых как P. gingivalis ПЦР методом. Для пересева смешанных культур и дальнейшего выделения
чистых культур P.gingivalis возможно использование бруцеллагара производства Оболенск (Россия) с добавлением чёрного
альбумина и менахинона (витамин К2
).
Annotation
Diagnosis of the etiological agents of chronic periodontitis, gram-negative anaerobic bacteria Porphyromonas gingivalis, is difficult
due to the high demands of this species for additional growth factors. The aim of the study was to increase the sensitivity of the
nutrient medium and reduce the time required to isolate a pure culture of P. gingivalis. Microbiological and molecular genetic methods
were used in the work. As a result, the Brucellagar nutrient medium was selected taking into account the cultural characteristics of
this periodontopathogen with the addition of black albumin and menaquinone (vitamin K2
). Small, round, mucous, dark-pigmented
colonies grew on the resulting nutrient medium, which were identified by PCR testing as P. gingivalis. Based on the results obtained,
for reseeding mixed cultures and further isolating pure cultures of P. gingivalis, it is possible to use brucellagar produced in Obolensk
(Russia) with the addition of black albumin and menaquinone (vitamin K2
).
Key words: Porphyromonas gingivalis; culture methods; black albumin and menaquinone
Список литературы
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Modern methods of diagnosis of periodontal diseases: opportunities
and prospects (review of literature). Klinicheskaya Laboratornaya Diagnostika. 2023; 68(9): 570-7. DOI: 10.51620/0869-2084-2023-68-9-
570-577. (in Russian)
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10.1038/nrmicro2873.
4. Xu W., Zhou W., Wang H., Liang S. Roles of Porphyromonas gingivalis and its virulence factors in periodontitis. Adv. Protein ChemStruct.
Biol. 2020; 120: 45-84. DOI: 10.1016/bs.apcsb.2019.12.001.
5. Chen C., Hemme C., Beleno J., Shi Z.J., Ning D., Qin Y., Tu Q., Jorgensen M., He Z., Wu L. et al. Oral microbiota of periodontal health
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ISME J. 2018; 12: 1210–24. DOI: 10.1038/s41396-017-0037-1.
6. Lamont R.J., Koo H., Hajishengallis G. The Oral microbiota: dynamic
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59. DOI: 10.1038/s41579-018-0089-x.
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inflammation and promote dysbiosis. Cell Host. Microbe. 2014; 15(6):
768-78. DOI: 10.1016/j.chom.2014.05.012.
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for in vitro cultivation of uncultured oral bacteria. J. Oral Maxillofac.
Pathol. 2021; 25(2): 266-71. DOI: 10.4103/0973-029X.325125.
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retrospective surveillance study. Journal of Clinical Periodontology.
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identification of Porphyromonas gingivalis validated with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Microb. Pathog. 2016; 94: 112-6. DOI: 10.1016/j.micpath.2016.01.021.
17. Roberts A., Matthews J.B., Socransky S.S., Freestone P.P.E., Williams
P.H., Chapple I.L.C. Stress and the periodontal diseases: effects of catecholamines on the growth of periodontal bacteria in vitro. Oral microbiology and immunology. 2002; 17 (5): 296-303. DOI: 10.1034/j.1399-
302X.2002.170506.x.
18. Ingalagi P., Bhat K.G., Kulkarni R.D., Kotrashetti V.S., Kumbar V.,
Kugaji M. Detection and comparison of prevalence of Porphyromonas
gingivalis through culture and Real Time-polymerase chain reaction in
subgingival plaque samples of chronic periodontitis and healthy individuals. J. Oral. Maxillofac. Pathol. 2022; 26(2): 288. DOI: 10.4103/
jomfp.jomfp_163_21.