Real-time PCR assay development for the control of vaccine against hemorrhagic fever with renal syndrome
- 作者: Egorova M.S.1, Kurashova S.S.1, Ishmukhametov A.A.1,2, Balovneva M.V.1, Deviatkin A.A.3, Safonova M.V.4, Ozherelkov S.V.1, Khapchaev Y.K.1, Balkina A.S.1, Belyakova A.V.1, Dzagurova T.K.1, Tkachenko EA.1
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隶属关系:
- FSBSI M.I. Chumakov Federal Scientific Center for Research and Development of Immune-and-Biological Products of Russian Academy of Sciences
- Institute for Translational Medicine and Biotechnology, FSAEI HE I.M. Sechenov First Moscow State Medical University of the Ministry of the Health of Russia
- Institute of Molecular Medicine, FSAEI HE I.M. Sechenov First Moscow State Medical University of the Ministry of the Health of Russia
- FSHI Plague Control Center» of the Federal Service for Surveillance of Consumer Rights Protection and Human Wellbeing (Rospotrebnadzor)
- 期: 卷 66, 编号 1 (2021)
- 页面: 65-73
- 栏目: ORIGINAL RESEARCH
- URL: https://ogarev-online.ru/0507-4088/article/view/118178
- DOI: https://doi.org/10.36233/0507-4088-30
- ID: 118178
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Introduction. Hemorrhagic fever with renal syndrome (HFRS) holds a leading place among natural focal human diseases in Russian Federation. There is no etiotropic therapy for the disease now. The vaccine prophylaxis is the most effective method to control this infection. The main criteria for inactivated vaccines evaluation are its immunogenicity and specific activity.
The study purposes were to develop a sensitive and specific real-time PCR method for viral RNA quantification in the inactivated vaccine and to study the correlation between the viral RNA amount and vaccine immunogenicity.
Material and methods. L-segment fragments of the Puumala, Hantaan, and Sochi vaccine strains were selected as diagnostic targets for oligonucleotides and fluorescent probes. The immunogenicity of experimental vaccines was determined by the induction of neutralizing antibodies in BALB/c mice.
Results. A highly specific, sensitive and reproducible real-time PCR method has been developed. The analytical sensitivity was 1.24 ± 1.5 x 102 copies/ml for Puumala virus; 1.16 ± 1.4 * 102 copies/ml for Hantaan; 1.32 ± 1.8 * 102 copies/ ml for Sochi, with a virus content of 1.5 ± 0.5 lg FFU/ml; 1.8 ± 0.5 lg FFU/ml and 2.2 ± 0.5 lg FFU/ml, respectively. The viral RNA amount in experimental vaccine preparations inactivated with β-propiolactone was proportional to the neutralizing antibodies titer observed in mice following the immunization.
Discussion. It was found that different virus inactivators differently affects the detected viral RNA amount, but not the vaccine immunogenicity, which indicates the same degree of the immunogenic proteins damage. The direct relationship between the viral RNA copy number and vaccine immunogenicity makes it possible to use this criterion for vaccine dosage preparation.
Conclusion. The developed method for viral RNA quantification is a promising tool for the specific activity control of the HFRS vaccine.
作者简介
M. Egorova
FSBSI M.I. Chumakov Federal Scientific Center for Research and Development of Immune-and-Biological Products of Russian Academy of Sciences
编辑信件的主要联系方式.
Email: masha_0787@mail.ru
ORCID iD: 0000-0002-6416-2573
Maria S. Egorova - Ph.D. (Biol.), Senior Researcher of the Laboratory of Hemorrhagic Fevers.
108819, Moscow
俄罗斯联邦S. Kurashova
FSBSI M.I. Chumakov Federal Scientific Center for Research and Development of Immune-and-Biological Products of Russian Academy of Sciences
Email: svetlanak886@yandex.ru
ORCID iD: 0000-0001-9934-699X
Kurashova Svetlana Sergeevna - Researcher, Laboratory of Hemorrhagic Fevers.
108819, Moscow
俄罗斯联邦A. Ishmukhametov
FSBSI M.I. Chumakov Federal Scientific Center for Research and Development of Immune-and-Biological Products of Russian Academy of Sciences; Institute for Translational Medicine and Biotechnology, FSAEI HE I.M. Sechenov First Moscow State Medical University of the Ministry of the Health of Russia
Email: aishmukhametov@gmail.com
ORCID iD: 0000-0001-6130-4145
Ishmukhametov Aidar Airatovich - PhD, MD, Professor, Corresponding Member of the Russian Academy of Sciences.
108819, Moscow, 119991, Moscow
俄罗斯联邦
M. Balovneva
FSBSI M.I. Chumakov Federal Scientific Center for Research and Development of Immune-and-Biological Products of Russian Academy of Sciences
Email: mashasm@yandex.ru
ORCID iD: 0000-0003-2198-7521
Balovneva Maria Vladimirovna - PhD, Leading Researcher, Laboratory of Hemorrhagic Fevers.
108819, Moscow
A. Deviatkin
Institute of Molecular Medicine, FSAEI HE I.M. Sechenov First Moscow State Medical University of the Ministry of the Health of Russia
Email: andreideviatkin@gmail.com
ORCID iD: 0000-0003-0789-4601
Devyatkin Andrey Andreevich - PhD, Leading Researcher, Laboratory of Molecular Biology and Biochemistry, Institute of Molecular Medicine.
119991, Moscow
俄罗斯联邦M. Safonova
FSHI Plague Control Center» of the Federal Service for Surveillance of Consumer Rights Protection and Human Wellbeing (Rospotrebnadzor)
Email: Marina-iris@mail.ru
ORCID iD: 0000-0002-5579-5694
Safonova Marina Viktorovna - Researcher, laboratory for viral infections I-II pathogenicity groups diagnostics of the Federal Public Health Institution "Anti-Plague Center".
119121, Moscow
俄罗斯联邦S. Ozherelkov
FSBSI M.I. Chumakov Federal Scientific Center for Research and Development of Immune-and-Biological Products of Russian Academy of Sciences
Email: ozherelkov@yandex.ru
ORCID iD: 0000-0001-9778-0485
Ozherelkov Sergey Viktorovich – PhD, DSc, Leading Researcher, tick-borne encephalitis and other viral encephalitis laboratory.
108819, Moscow
俄罗斯联邦Yu. Khapchaev
FSBSI M.I. Chumakov Federal Scientific Center for Research and Development of Immune-and-Biological Products of Russian Academy of Sciences
Email: hapchaev_uh@chumakovs.su
ORCID iD: 0000-0003-1613-5228
Khapchaev Yusuf Khadzhibekovich - PhD, DSc, Head of the Polio Vaccine Department.
108819, Moscow
俄罗斯联邦A. Balkina
FSBSI M.I. Chumakov Federal Scientific Center for Research and Development of Immune-and-Biological Products of Russian Academy of Sciences
Email: balkina_as@chumakovs.su
ORCID iD: 0000-0002-9704-7774
Balkina Alexandra Sergeevna - Researcher, Laboratory of Hemorrhagic Fevers.
108819, Moscow
俄罗斯联邦A. Belyakova
FSBSI M.I. Chumakov Federal Scientific Center for Research and Development of Immune-and-Biological Products of Russian Academy of Sciences
Email: belyakova_av@chumakovs.su
Belyakova Alla Vladimirovna - PhD, Scientific Secretary.
108819, Moscow
俄罗斯联邦T. Dzagurova
FSBSI M.I. Chumakov Federal Scientific Center for Research and Development of Immune-and-Biological Products of Russian Academy of Sciences
Email: centrglps@yandex.ru
ORCID iD: 0000-0002-6656-1682
Dzagurova Tamara Kazbekovna - PhD, MD, Leading Researcher, Head of the Laboratory of Hemorrhagic Fevers.
108819, Moscow
俄罗斯联邦E Tkachenko
FSBSI M.I. Chumakov Federal Scientific Center for Research and Development of Immune-and-Biological Products of Russian Academy of Sciences
Email: evgeniytkach@mail.ru
ORCID iD: 0000-0002-6829-1241
Tkachenko Evgeniy Aleksandrovich - PhD, MD, Professor, Scientific Superviser.
108819, Moscow
俄罗斯联邦参考
- Kruger D.H., Figueiredo L.T., Song J.W., Klempa B. Hantaviruses -globally emerging pathogens. J. Clin. Virol. 2014; 64: 128-36. https://doi.org/10.1016/jjcv.2014.08.033.
- Lahdevirta J. Nephropathia epidemica in Finland. A clinical, histological and epidemiological study. Ann. Clin. Res. 1971; 3(Suppl. 8): 1-154.
- Schmaljohn C., Hjelle B. Hantaviruses: a global disease problem. Emerg. Infect. Dis. 1997; 3(2): 95-104. https://doi.org/10.3201/eid0302.970202.
- Maes P., Li S., Verbeeck J., Keyaerts E., Clement J., Van Ranst M. Evaluation of the efficacy of disinfectants against Puumala hantavirus by real-time RT-PCR. J. Virol. Methods. 2007; 141(1): 111-5. https://doi.org/10.1016/jjviromet.2006.11.037.
- Plyusnin A., Vapalahti O., Vaheri A. Hantaviruses: genome structure, expression and evolution. J. Gen. Virol. 1996; 77(Pt. 11): 2677-87. https://doi.org/10.1099/0022-1317-77-11-2677.
- Schmaljohn C.S., Dalrymple J.M. Analysis of Hantaan virus RNA: evidence for a new genus of Bunyaviridae. Virology. 1983; 131(2): 482-91. https://doi.org/10.1016/0042-6822(83)90514-7.
- Tkachenko E.A., Ishmukhametov A.A., Dzagurova T.K., Bern-shtein A.D., Morozov V.G., Sinyugina A.A., et al. Hemorrhagic fever with renal syndrome, Russia. Emerg. Infect. Dis. 2019; 25(12): 2325-8. https://doi.org/10.3201/eid2512.181649.
- Liu R., Ma H., Shu J., Zhang Q., Han M., Liu Z., et al. Vaccines and therapeutics against hantaviruses. Front. Microbiol. 2019; 10: 2989. https://doi.org/10.3389/fmicb.2019.02989.
- Бархалева О.А., Воробьева М.С., Ладыженская И.П., Ткаченко Е.А., Дзагурова Т.К. Вакцина против геморрагической лихорадки с почечным синдромом. БИОпрепараты. Профилактика, диагностика, лечение. 2011; (1): 27-30.
- Jiang W., Ma N., Hui Q., Chen B., Qiu Y., Sun H., et al. Type inactivated bivalent hantavirus vaccines using dual fluorescence quantitative real time PCR: Establishment and evaluation. J. Med. Virol. 2017; 89(1): 10-6. https://doi.org/10.1002/jmv.24599.
- Ткаченко Е.А., Ишмухаметов А.А., Дзагурова Т.К., Синюгина А.А., Коротина Н.А., Набатников П.А., и др. Разработка экспериментально-промышленной технологии производства вакцины для профилактики геморрагической лихорадки с почечным синдромом. Ремедиум. Журнал о российском рынке лекарств и медицинской технике. 2015; (6): 47-53.
- Курашова С.С., Ишмухаметов А.А., Егорова MG., Баловнева М.В., Дзагурова Т.К., Ткаченко Е.А. Сравнительная характеристика инактивирующих агентов для создания вакцины против геморрагической лихорадки с почечным синдромом. Эпидемиология и вакцинопрофилактика. 2018; 17(4): 26-8. https://doi.org/10.31631/2073-3046-2018-17-4-26-29.
- Епринцев А.Т., Попов В.Н., Федорин Д.Н. Идентификация и исследование экспрессии генов: Учебно-методическое пособие для вузов. Воронеж; 2008: 14-5.
- Дзагурова Т.К., Солопова О.Н., Свешников П.Г., Коротина Н.А., Баловнева М.В., Леонович О.А., и др. Разработка иммуноферментной тест-системы на основе моноклональных антител для определения специфической активности вакцины против геморрагической лихорадки с почечным синдромом. Вопросы вирусологии. 2013; 58(1): 40-4.
- Дзагурова Т.К., Ткаченко Е.А., Башкирцев В.Н., Окулова Н.М., Апекина Н.С., Коротина Н.А., и др. Выделение и идентификация штаммов хантавирусов - возбудителей ГЛПС в Европейской части России.Медицинская вирусология. 2008; 25: 142-50.
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