Журнал микробиологии, эпидемиологии и иммунобиологии. 2018; 1: 45-52
ВАРИАБЕЛЬНОСТЬ НУКЛЕОТИДНЫХ ПОСЛЕДОВАТЕЛЬНОСТЕЙ ГЕНА arp У РОССИЙСКИХ ИЗОЛЯТОВ TREPONEMA PALLIDUM
https://doi.org/10.36233/0372-9311-2018-3-45-52Аннотация
Цель. Анализ вариабельности нуклеотидных последовательностей внутреннего фрагмента гена arp у современных российских штаммов T. pallidum subsp. pallidum. Материалы и методы. В работе использованы 57 клинических изолятов, полученных в 2016 — 2017 гг. из специализированных медицинских учреждений дерматовенерологического профиля Центрального (Калужская область), Северо-Кавказского (Ставропольский край) и Сибирского (Республика Тыва) федеральных округов. Для исследования нуклеотидных последовательностей гена arp использована технология капиллярного секвенирования. Результаты. Предложена двухраундовая амплификация гена arp, обеспечивающая корректное прочтение его внутреннего участка. В составе данных участков описаны 4 варианта 60-нуклеотидных повторов, отличающихся по составу 6, 8 и 15 — 17 триплетов. Охарактеризованы различные комбинации подобных повторов, соответствующие референсному штамму Nichols, глобально распространенной геногруппе Street 14, а также и впервые обнаруженному региональному варианту Stavropol. Заключение. Продемонстрирована целесообразность секвенирования гена arp как инструмента повышения эффективности молекулярного типирования T. pallidum.
Список литературы
1. Белозоров А.П., Сокол О.А. Молекулярное типирование микроорганизма Treponema pallidum подвид pallidum. Дерматологiя та венерологiя. 2013, 4 (62): 5-11.
2. Кубанов А.А., Воробьев Д.В., Обухов А.П., Образцова О.А., Дерябин Д.Г. Молекулярная эпидемиология Treponema pallidum в приграничном регионе Российской Федерации (Республика Тыва). Молекулярная генетика, микробиология и вирусология. 2017, 1: 30-34.
3. Cejková D., Zobaníková M., Chen L. et al. Whole genome sequences of three Treponema pallidum ssp. pertenue strains: yaws and syphilis treponemes differ in less than 0.2% of the genome sequence. PLoS Negl. Trop. Dis. 2012, 6(1). doi: 10.1371.
4. Dekker M.C., van Duijn C.M. Prospects of genetic epidemiology in the 21st century. Eur. J. Epidemiol. 2003, 18 (7): 607-616.
5. Flasarová M., Pospíšilová P., Mikalová L. et al. Sequencing-based molecular typing of Treponema pallidum strains in the Czech Republic: all identified genotypes are related to the sequence of the SS14 strain. Acta Derm. Venereol. 2012, 92: 669-674.
6. Fraser C.M., Norris S.J., Weinstock G.M. et al. Complete genome sequence of Treponema pallidum, the syphilis spirochete. Science. 1998, 281 (5375): 375-388.
7. Harper K.N., Liu H., Ocampo P.S. et al. The sequence of the acidic repeat protein (arp) gene differentiates venereal from nonvenereal Treponema pallidum subspecies, and the gene has evolved under strong positive selection in the subspecies that causes syphilis. FEMS Immunol. Med. Microbiol. 2008, 53 (3): 322-332.
8. Liu H., Rodes B., Chen C.Y., Steiner B. New tests for syphilis: rational design of a PCR method for detection of Treponema pallidum in clinical specimens using unique regions of the DNA polymerase I gene. J. Clin. Microbiol. 2001, 39 (5): 1941-1946.
9. Liu H., Rodes B., George R., Steiner B. Molecular characterization and analysis of a gene encoding the acidic repeat protein (Arp) of Treponema pallidum. J. Med. Microbiol. 2007, 56 (6): 715-721.
10. Ma D.Y., Giacani L., Centurión-Lara A. The molecular epidemiology of Treponema pallidum subspecies pallidum. Sex Health. 2015, 12 (2): 141-147.
11. Marra C.M., Sahi S.K., Tantalo L.C. et al. Enhanced molecular typing of Treponema pallidum: geographical distribution of strain types and association with neurosyphilis. J. Infect. Dis. 2010, 202: 1380-1388.
12. Matejkova P., Strouhal M., Smajs D. et al. Complete genome sequence of Treponema pallidum spp. pallidum strain SS14 determined with oligonucleotide arrays. BMC Microbiol, 2008, 8:76. doi:10.1186/1471-2180-8-76.
13. Pillay A., Liu H., Chen C.Y. et al. Molecular subtyping of Treponema pallidum subspecies pallidum. Sex. Transm. Dis. 1998, 25 (8): 408-414.
Journal of microbiology, epidemiology and immunobiology. 2018; 1: 45-52
Arp GENE NUCLEOTID SEQUENCES VARIABILITY IN RUSSIAN TREPONEMA PALLIDUM ISOLATES
https://doi.org/10.36233/0372-9311-2018-3-45-52Abstract
Aim. Analysis of the arp gene internal fragment nucleotide sequences variability in modern russian T. pallidum subsp. pallidum strains. Materials and methods. 57 T. pallidum isolates obtained from specialized dermatovenerologic clinics of the Central (Kaluga), the North Caucasus (Stavropol) and the Siberian (Tyva) regions in 2016 — 2017 were used in the study. The sequensing of the arp gene was performed using capillary electrophoresis technology. Results. A two-round amplification of the arp gene have been proposed, which ensures a correct reading of its internal region. Four variants of 60-nucleotide repeats in the internal arp fragment are described, which differing in 6, 8 and 15 — 17 codons compositions. Various combinations of these repeats, corresponding to the reference Nichols strain, globally distributed Street 14 genogroup, and the firstly described Stavropol regional variant are shown. Conclusion. The prospect of arp gene sequencing as a way to increase T. pallidum molecular typing efficiency is postulated.
References
1. Belozorov A.P., Sokol O.A. Molekulyarnoe tipirovanie mikroorganizma Treponema pallidum podvid pallidum. Dermatologiya ta venerologiya. 2013, 4 (62): 5-11.
2. Kubanov A.A., Vorob'ev D.V., Obukhov A.P., Obraztsova O.A., Deryabin D.G. Molekulyarnaya epidemiologiya Treponema pallidum v prigranichnom regione Rossiiskoi Federatsii (Respublika Tyva). Molekulyarnaya genetika, mikrobiologiya i virusologiya. 2017, 1: 30-34.
3. Cejková D., Zobaníková M., Chen L. et al. Whole genome sequences of three Treponema pallidum ssp. pertenue strains: yaws and syphilis treponemes differ in less than 0.2% of the genome sequence. PLoS Negl. Trop. Dis. 2012, 6(1). doi: 10.1371.
4. Dekker M.C., van Duijn C.M. Prospects of genetic epidemiology in the 21st century. Eur. J. Epidemiol. 2003, 18 (7): 607-616.
5. Flasarová M., Pospíšilová P., Mikalová L. et al. Sequencing-based molecular typing of Treponema pallidum strains in the Czech Republic: all identified genotypes are related to the sequence of the SS14 strain. Acta Derm. Venereol. 2012, 92: 669-674.
6. Fraser C.M., Norris S.J., Weinstock G.M. et al. Complete genome sequence of Treponema pallidum, the syphilis spirochete. Science. 1998, 281 (5375): 375-388.
7. Harper K.N., Liu H., Ocampo P.S. et al. The sequence of the acidic repeat protein (arp) gene differentiates venereal from nonvenereal Treponema pallidum subspecies, and the gene has evolved under strong positive selection in the subspecies that causes syphilis. FEMS Immunol. Med. Microbiol. 2008, 53 (3): 322-332.
8. Liu H., Rodes B., Chen C.Y., Steiner B. New tests for syphilis: rational design of a PCR method for detection of Treponema pallidum in clinical specimens using unique regions of the DNA polymerase I gene. J. Clin. Microbiol. 2001, 39 (5): 1941-1946.
9. Liu H., Rodes B., George R., Steiner B. Molecular characterization and analysis of a gene encoding the acidic repeat protein (Arp) of Treponema pallidum. J. Med. Microbiol. 2007, 56 (6): 715-721.
10. Ma D.Y., Giacani L., Centurión-Lara A. The molecular epidemiology of Treponema pallidum subspecies pallidum. Sex Health. 2015, 12 (2): 141-147.
11. Marra C.M., Sahi S.K., Tantalo L.C. et al. Enhanced molecular typing of Treponema pallidum: geographical distribution of strain types and association with neurosyphilis. J. Infect. Dis. 2010, 202: 1380-1388.
12. Matejkova P., Strouhal M., Smajs D. et al. Complete genome sequence of Treponema pallidum spp. pallidum strain SS14 determined with oligonucleotide arrays. BMC Microbiol, 2008, 8:76. doi:10.1186/1471-2180-8-76.
13. Pillay A., Liu H., Chen C.Y. et al. Molecular subtyping of Treponema pallidum subspecies pallidum. Sex. Transm. Dis. 1998, 25 (8): 408-414.
