zdorПроблемы здоровья и экологииHealth and Ecology Issues2220-09672708-6011Gomel State Medical University10.51523/2708-6011.2022-19-1-13zdor-2182Research ArticleЭКСПЕРИМЕНТАЛЬНАЯ МЕДИЦИНА И БИОЛОГИЯEXPERIMENTAL MEDICINE AND BIOLOGYСравнение различных вариантов выявления ДНК вирусов TTV, TTMDV и TTMVComparison of different DNA detection options for TTV, TTMDV, and TTMV viruseshttps://orcid.org/0000-0002-1931-4224ОсипкинаО. В.OsipkinaO. V.

Осипкина Ольга Викторовна, заведующий НИЛ

г. Гомель

Olga V. Osipkina, Head of the Research Laboratory

Gomel

olga.osipkina@mail.ruhttps://orcid.org/0000-0002-9435-6109ВоропаевЕ. В.VoropaevE. V.

Воропаев Евгений Викторович, кандидат медицинских наук, доцент, проректор по научной работе

г. Гомель

Evgenii V. Voropaev, PhD (Med), Associate Professor, Vice-Rector in charge of scientific work

Gomel

voropaev.evgenii@gmail.comhttps://orcid.org/0000-0002-0449-5026МицураВ. М.MitsuraV. M.

Мицура Виктор Михайлович, доктор медицинских наук, доцент, заместитель директора по научной работе, ГУ «Республиканский научно-практический центр радиационной медицины и экологии человека»; профессор кафедры инфекционных болезней, УО «Гомельский государственный медицинский университет»

Гомель

Viktor M. Mitsura, DMedSc, Associate Professor, Deputy Director for Research, Republican Research Center for Radiation Medicine and Human Ecology; Professor at the Department of Infectious Diseases, Gomel State Medical University

Gomel

mitsura_victor@tut.byhttps://orcid.org/0000-0003-1974-5355ТерешковД. В.TereshkovD. V.

Терешков Дмитрий Валерьевич, заведующий инфекционным отделением

г. Гомель

Dmitriy V. Tereshkov, Head of the Infectious Diseases Department

Gomel

tereshkovd@tut.byhttps://orcid.org/0000-0001-9148-487XКовалёвА. А.KovalevA. A.

Ковалёв Алексей Алексеевич, инженер-программист отдела науки и научно-методической информации

г. Гомель

Alexey A. Kovalev, software engineer of the Department of Science and Scientific and Methodical Information

Gomel

kovalev.data.analysis.gsmu@yandex.byГомельский государственный медицинский университетGomel State Medical UniversityГомельский государственный медицинский университет; Республиканский научно-практический центр радиационной медицины и экологии человекаGomel State Medical University; Republican Research Center for Radiation Medicine and Human EcologyГомельская областная инфекционная клиническая больницаGomel Regional Clinical Hospital of Infectious Diseases202230032022191102108Copyright © Осипкина О.В., Воропаев Е.В., Мицура В.М., Терешков Д.В., Ковалёв А.А., 20222022Осипкина О.В., Воропаев Е.В., Мицура В.М., Терешков Д.В., Ковалёв А.А.Osipkina O.V., Voropaev E.V., Mitsura V.M., Tereshkov D.V., Kovalev A.A.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://journal.gsmu.by/jour/article/view/2182Цель исследования

Цель исследования. Сравнить различные варианты выявления ДНК TTV, TTMDV и TTMV в плазме крови пациентов с различными заболеваниями печени и у лиц без признаков заболеваний печени.

Материалы и методы

Материалы и методы. Для выявления ДНК TTV, TTMDV и TTMV использовался метод ПЦР.

Результаты

Результаты. Обнаружена достаточно высокая частота выявления ДНК вирусов TTV при использовании разных лабораторных подходов. Частота выявления ДНК TTV значимо выше при использовании праймеров для некодирующего региона UTR — 77,3 % при сравнении с кодирующим ORF1 — 38,4 % (р < 0,001) и коммерческим набором реагентов — 53 % (р < 0,005).

Заключение

Заключение. У пациентов с заболеваниями печени по сравнению со здоровыми лицами значимо чаще выявляются ДНК TTV (90,3 и 65,6 % по UTR-региону и с использованием коммерческого набора соответственно), ДНК TTMV (83,9 % — UTR-регион) и микст ДНК вирусов TTV + TTMDV +TTMV (62,4 % — UTR-регион). С целью стандартизации определения ДНК вирусов семейства TTV методом ПЦР целесообразно создание панели сывороток, содержащей достоверно положительные и отрицательные образцы.

Objective

Objective. To compare different variants of TTV, TTMDV, and TTMV DNA detection in the blood plasma of patients with various liver diseases and in individuals without signs of liver disease.

Materials and methods

Materials and methods. To detect TTV TTMDV, and TTMV DNA, the PCR method was used.

Results

Results. A fairly high frequency of TTV virus DNA detection was found using different laboratory approaches. The frequency of TTV DNA detection was significantly highest when using the primers for the non-coding region UTR – 77.3 % compared with the coding region ORF1 – 38.4 % (p < 0.001) and the commercial kit 53 % (p < 0.005).

Conclusion

Conclusion. TTV DNA is detected significantly more often in patients with liver diseases compared with healthy individuals (90.3 % and 65.6 % in the UTR region and using the commercial kit, respectively), TTMV DNA (83.9 % – UTR region) and mixed DNA of the TTV + TTMDV + TTMV viruses (62.4 % – UTR region). To standardize the DNA detection of the TTV family viruses by the PCR method, it is advisable to create a panel of sera containing reliably positive and negative samples.

TTVПЦРлабораторная диагностикаTTVPCRlaboratory diagnosticsИсследование проведено в рамках выполнения НИОК(Т)Р «Разработать методы диагностики и патогенетического лечения хронических прогрессирующих заболеваний паренхиматозных органов и связанных с ними состояний, сопровождающихся нарушением процессов регенерации», № госрегистрации 20190387 от 29.03.2019 г.The study was carried out within the framework of the R&D project “To develop methods for the diagnosis and pathogenetic treatment of chronic progressive diseases of the parenchymal organs and related conditions accompanied by failed regeneration processes” under State Registration No.20190387 dated 29.03.2019.ReferencesKowarsky M, Camunas-Soler J, Kertesz M, De Vlaminck I, Koh W, Pan W, et al. Numerous uncharacterized and highly divergent microbes which colonize humans are revealed by circulating cell-free DNA. Proceedings of the National Academy of Sciences of the United States of America. 2017;114(36):9623-9628. https://doi.org/10.1073/pnas.1707009114Kowarsky M, Camunas-Soler J, Kertesz M, De Vlaminck I, Koh W, Pan W, et al. Numerous uncharacterized and highly divergent microbes which colonize humans are revealed by circulating cell-free DNA. Proceedings of the National Academy of Sciences of the United States of America. 2017;114(36):9623-9628. https://doi.org/10.1073/pnas.1707009114Nishizawa T, Okamoto H, Konishi K, Yoshizawa H, Miyakawa Y, Mayumi M. A novel DNA virus (TTV) associated with elevated transaminase levels in posttransfusion hepatitis of unknown etiology. Biochem Biophys Res Commun. 1997;241(1):92-97. https://doi.org/10.1006/bbrc.1997.7765Nishizawa T, Okamoto H, Konishi K, Yoshizawa H, Miyakawa Y, Mayumi M. A novel DNA virus (TTV) associated with elevated transaminase levels in posttransfusion hepatitis of unknown etiology. Biochem Biophys Res Commun. 1997;241(1):92-97. https://doi.org/10.1006/bbrc.1997.7765Okamoto H. History of discoveries and pathogenicity of TT viruses. Curr Top Microbiol Immunol. 2009;331:1-20. https://doi.org/10.1007/978-3-540-70972-5_1Okamoto H. History of discoveries and pathogenicity of TT viruses. Curr Top Microbiol Immunol. 2009;331:1-20. https://doi.org/10.1007/978-3-540-70972-5_1Miyata H, Tsunoda H, Kazi A, Yamada A, Khan MA, Murakami J, Kamahora T, Shiraki K, Hino S. Identification of a novel GC-rich 113-nucleotide region to complete the circular, single-stranded DNA genome of TT virus, the first human circovirus. J Virol. 1999;73(5):3582-3586. https://doi.org/10.1128/JVI.73.5.3582-3586.1999Miyata H, Tsunoda H, Kazi A, Yamada A, Khan MA, Murakami J, Kamahora T, Shiraki K, Hino S. Identification of a novel GC-rich 113-nucleotide region to complete the circular, single-stranded DNA genome of TT virus, the first human circovirus. J Virol. 1999;73(5):3582-3586. https://doi.org/10.1128/JVI.73.5.3582-3586.1999Virus Taxonomy: 2020 Release [Electronic resource]. International Committee on Taxonomy of Viruses [data of access 23 January 2022]. Available from: https://https://talk.ictvonline.org/taxonomy/Virus Taxonomy: 2020 Release [Electronic resource]. International Committee on Taxonomy of Viruses [data of access 23 January 2022]. Available from: https://talk.ictvonline.org/taxonomy/Reshetnyak V, Maev I, Burmistrov A, Chekmazov I, Karlovich T. Torque teno virus in liver diseases: On the way towards unity of view. World J Gastroenterol. 2020;26(15):1691-1707. https://doi.org/10.3748/wjg.v26.i15.1691Reshetnyak V, Maev I, Burmistrov A, Chekmazov I, Karlovich T. Torque teno virus in liver diseases: On the way towards unity of view. World J Gastroenterol. 2020;26(15):1691-1707. https://doi.org/10.3748/wjg.v26.i15.1691Spandole S, Cimponeriu D, Berca L, Mihăescu G. Human anelloviruses: an update of molecular, epidemiological and clinical aspects. Arch Virol. 2015;160:893-908.Spandole S, Cimponeriu D, Berca L, Mihăescu Human anelloviruses: an update of molecular, epidemiological and clinical aspects. Arch Virol 2015;160:893-908.Maggi F, Bendinelli M. Immunobiology of the Torque teno viruses and other anelloviruses. Curr Top Microbiol Immunol. 2009;331:65-90. https://doi.org/10.1007/978-3-540-70972-5_5Maggi F, Bendinelli M. Immunobiology of the Torque teno viruses and other anelloviruses. Curr Top Microbiol Immunol. 2009;331:65-90. https://doi.org/10.1007/978-3-540-70972-5_5Schmidt L, Jensen B, Walker A, et al. Torque Teno Virus plasma level as novel biomarker of retained immunocompetence in HIV-infected patients. Infection 2021;49:501-509. https://doi.org/10.1007/s15010-020-01573-7Schmidt L, Jensen B, Walker A, et al. Torque Teno Virus plasma level as novel biomarker of retained immunocompetence in HIV-infected patients. Infection 2021;49:501-509. https://doi.org/10.1007/s15010-020-01573-7Chen T, Väisänen E, Mattila P, Hedman K, Söderlund-Venermo M. Antigenic diversity and seroprevalences of torque teno viruses in children and adults by ORF2-based immunoassays. J Gen Virol. 2013;94:409-417. https://doi.org/10.1099/vir.0.046862-0Chen T, Väisänen E, Mattila P, Hedman K, Söderlund-Venermo M. Antigenic diversity and seroprevalences of torque teno viruses in children and adults by ORF2based immunoassays. J Gen Virol. 2013;94:409-417. https://doi.org/10.1099/vir.0.046862-0Mankotia D, Irshad M. Development of an Immunoassay for Detection of Torque Teno Virus (TTV) Antibodies Using the N22 Expression Product from TTV Genotype 2. Intervirology. 2017;60(5):207-216. https://doi.org/10.1159/000487481Mankotia D, Irshad M. Development of an Immunoassay for Detection of Torque Teno Virus (TTV) Antibodies Using the N22 Expression Product from TTV Genotype 2. Intervirology. 2017;60(5):207-216. https://doi.org/10.1159/000487481Осипкина ОВ, Воропаев ЕВ, Мицура ВМ, Терешков ДВ, Зятьков АА, Баранов ОЮ. Метод молекулярной диагностики ТТ-вирусной инфекции: инструкция по применению. Гомель; 2018. 15 с.Osipkina OV, Voropaev EV, Mitsura VM, Tereshkov DV, Zyatskov AA, Baranov OYu. Method for molecular diagnostics of TT virus infection: instructions for use. Gomel; 2018. 15 p. (in Russ.).Ninomiya M, Takahashi M, Nishizawa T, Shimosegawa T, Okamoto H. Development of PCR Assays with Nested Primers Specific for Differential Detection of Three Human Anelloviruses and Early Acquisition of Dual or Tri- ple Infection during Infancy. Journal of Clinical Microbiology. 2008;2(46):507-514. https://doi.org/10.1128/JCM.01703-07Ninomiya M, Takahashi M, Nishizawa T, Shimosegawa T, Okamoto H. Development of PCR Assays with Nested Primers Specific for Differential Detection of Three Human Anelloviruses and Early Acquisition of Dual or Tri- ple Infection during Infancy. Journal of Clinical Microbiology. 2008;2(46):507-514. https://doi.org/10.1128/JCM.01703-07Hu Y, Al-Moslih M, Al Ali M, Khameneh S, Perkins H, Diaz-Mitoma F, et al. Molecular detection method for all known genotypes of TT virus (TTV) and TTV-like viruses in thalassemia patients and healthy individuals. Journal of clinical microbiology. 2005;43(8):3747-3754. https://doi.org/10.1128/JCM.43.8.3747-3754.2005Hu Y, Al-Moslih M, Al Ali M, Khameneh S, Perkins H, Diaz-Mitoma F, et al. Molecular detection method for all known genotypes of TT virus (TTV) and TTV-like viruses in thalassemia patients and healthy individuals. Journal of clinical microbiology. 2005;43(8):3747-3754. https://doi.org/10.1128/JCM.43.8.3747-3754.2005

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