Статья

Antigenic properties of sARs-CoV-2/human/RUs/nsk-FRCFtM-1/2020 coronavirus isolate from a patient in novosibirsk [аНтиГеННые СвойСтва изолята короНавируСа SARS-COV-2/humAn/RuS/nSk-FRCFTm-1/2020, выделеННоГо от ПациеНта в НовоСибирСке]

A. Chepurnov, K. Sharshov, E. Kazachinskaya, Y. Kononova, E. Kazachkova, O. Khripko, K. Yurchenko, A. Alekseev, M. Voevoda, A. Shestopalov,
2021

Objective: isolation of coronavirus SARS-CoV-2 from clinical sample of patient with COVID-19 in Novosibirsk; obtaining a purified and inactivated viral antigen and study of its antigenic properties. Materials and methods: virus isolation was carried out in Vero cell culture from nasopharyngeal swab positive on SARS-CoV-2 RNA. The efficiency of SARS-CoV-2 replication in cell culture was assessed on the appearance of cytopathic effect (CPE) and the presence of viral RNA in cultural medium with reverse transcription – polymerase chain reaction (RT-PCR). Purification, concentration and inactivation of the viral preparation were carried out according to standard methods. The purity of the purified preparation and the profile of viral proteins were determined by electrophoresis in 10% polyacrylamide gel (PAG) with the addition of sodium dodecyl sulfate (SDS). The presence and specificity of viral proteins were detected using COVID-19 convalescent’s sera with enzyme-linked immunosorbent assay (ELISA) and immunoblotting. Results: SARS-CoV-2/human/ RUS/Nsk-FRCFTM-1/2020 isolate was obtained after passage on Vero cells from a virus-containing clinical sample. A purified, concentrated, inactivated, whole-virion antigen was obtained. It contains three structural proteins: glycoprotein S (approximately 200 kDa), nucleoprotein N (48 kDa), and matrix protein M (20-25 kDa). All viral proteins were detected with serum antibodies of COVID-19 convalescents. Conclusion: SARS-CoV-2 coronavirus can be isolated in Vero cell culture. The antigenic specificity of the three structural viral proteins (S, N, and M) is preserved in the purified inactivated viral preparation. The inactivated whole-virion antigen of SARS-CoV-2/human/RUS/Nsk-FRCFTM-1/2020 isolate can be used to study the antigenic immunomodulating properties of viral proteins, to obtain immune sera of laboratory animals, and also as a component of test systems for the detection of specific antibodies with ELISA and immunoblotting. © 2020 Interregional public organization Association of infectious disease specialists of Saint-Petersburg and Leningrad region (IPO AIDSSPbR). All rights reserved.

Цитирование

Похожие публикации

Документы

Источник

Версии

  • 1. Version of Record от 2021-04-27

Метаданные

Об авторах
  • A. Chepurnov
    Federal Research Center of Fundamental and Translational Medicine, Siberian Branch of Russian Academy of Sciences, Novosibirsk, Russian Federation
  • K. Sharshov
  • E. Kazachinskaya
  • Y. Kononova
  • E. Kazachkova
  • O. Khripko
  • K. Yurchenko
  • A. Alekseev
  • M. Voevoda
  • A. Shestopalov
Название журнала
  • Jurnal Infektologii
Том
  • 12
Выпуск
  • 3
Страницы
  • 42-50
Ключевые слова
  • glycoprotein s; matrix protein m; nucleoprotein n; structural protein; unclassified drug; viral protein; virus antigen; virus RNA; antigen specificity; antigenicity; Article; coronavirus disease 2019; cytopathogenic effect; electrophoresis; enzyme linked immunosorbent assay; human; immunoblotting; reverse transcription polymerase chain reaction; Severe acute respiratory syndrome coronavirus 2; throat culture; Vero cell line; virus inactivation; virus isolation; virus purification; virus replication
Издатель
  • Interregional public organization Association of infectious disease specialists of Saint-Petersburg and Leningrad region (IPO AIDSSPbR)
Тип документа
  • journal article
Тип лицензии Creative Commons
  • CC
Правовой статус документа
  • Свободная лицензия
Источник
  • scopus