Antigenic properties of sARs-CoV-2/human/RUs/nsk-FRCFtM-1/2020 coronavirus isolate from a patient in novosibirsk
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...
| Main Authors: | , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | Russian |
| Published: |
Journal Infectology
2020-08-01
|
| Series: | Журнал инфектологии |
| Subjects: | |
| Online Access: | https://journal.niidi.ru/jofin/article/view/1077 |
| _version_ | 1797702652705898496 |
|---|---|
| author | A. A. Chepurnov K. A. Sharshov E. I. Kazachinskaya Yu. V. Kononova E. A. Kazachkova O. P. Khripko K. S. Yurchenko A. Yu. Alekseev M. I. Voevoda A. M. Shestopalov |
| author_facet | A. A. Chepurnov K. A. Sharshov E. I. Kazachinskaya Yu. V. Kononova E. A. Kazachkova O. P. Khripko K. S. Yurchenko A. Yu. Alekseev M. I. Voevoda A. M. Shestopalov |
| author_sort | A. A. Chepurnov |
| collection | DOAJ |
| description | 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 SARSCoV-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. |
| first_indexed | 2024-03-12T04:53:10Z |
| format | Article |
| id | doaj.art-bf76a594153c45649a1a405730d6265a |
| institution | Directory Open Access Journal |
| issn | 2072-6732 |
| language | Russian |
| last_indexed | 2024-03-12T04:53:10Z |
| publishDate | 2020-08-01 |
| publisher | Journal Infectology |
| record_format | Article |
| series | Журнал инфектологии |
| spelling | doaj.art-bf76a594153c45649a1a405730d6265a2023-09-03T09:18:32ZrusJournal InfectologyЖурнал инфектологии2072-67322020-08-01123425010.22625/2072-6732-2020-12-3-42-50830Antigenic properties of sARs-CoV-2/human/RUs/nsk-FRCFtM-1/2020 coronavirus isolate from a patient in novosibirskA. A. Chepurnov0K. A. Sharshov1E. I. Kazachinskaya2Yu. V. Kononova3E. A. Kazachkova4O. P. Khripko5K. S. Yurchenko6A. Yu. Alekseev7M. I. Voevoda8A. M. Shestopalov9Федеральный исследовательский центр фундаментальной и трансляционной медицины Сибирского отделения РАНФедеральный исследовательский центр фундаментальной и трансляционной медицины Сибирского отделения РАНФедеральный исследовательский центр фундаментальной и трансляционной медицины Сибирского отделения РАНФедеральный исследовательский центр фундаментальной и трансляционной медицины Сибирского отделения РАНФедеральный исследовательский центр фундаментальной и трансляционной медицины Сибирского отделения РАНФедеральный исследовательский центр фундаментальной и трансляционной медицины Сибирского отделения РАНФедеральный исследовательский центр фундаментальной и трансляционной медицины Сибирского отделения РАНФедеральный исследовательский центр фундаментальной и трансляционной медицины Сибирского отделения РАНФедеральный исследовательский центр фундаментальной и трансляционной медицины Сибирского отделения РАНФедеральный исследовательский центр фундаментальной и трансляционной медицины Сибирского отделения РАНObjective: 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 SARSCoV-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.https://journal.niidi.ru/jofin/article/view/1077коронавирус тяжелого острого респираторного синдрома 2 типа (sars-cov-2)коронавирусная инфекция 2019 (covid-19)культура клеток veroвирусные белки |
| spellingShingle | A. A. Chepurnov K. A. Sharshov E. I. Kazachinskaya Yu. V. Kononova E. A. Kazachkova O. P. Khripko K. S. Yurchenko A. Yu. Alekseev M. I. Voevoda A. M. Shestopalov Antigenic properties of sARs-CoV-2/human/RUs/nsk-FRCFtM-1/2020 coronavirus isolate from a patient in novosibirsk Журнал инфектологии коронавирус тяжелого острого респираторного синдрома 2 типа (sars-cov-2) коронавирусная инфекция 2019 (covid-19) культура клеток vero вирусные белки |
| title | Antigenic properties of sARs-CoV-2/human/RUs/nsk-FRCFtM-1/2020 coronavirus isolate from a patient in novosibirsk |
| title_full | Antigenic properties of sARs-CoV-2/human/RUs/nsk-FRCFtM-1/2020 coronavirus isolate from a patient in novosibirsk |
| title_fullStr | Antigenic properties of sARs-CoV-2/human/RUs/nsk-FRCFtM-1/2020 coronavirus isolate from a patient in novosibirsk |
| title_full_unstemmed | Antigenic properties of sARs-CoV-2/human/RUs/nsk-FRCFtM-1/2020 coronavirus isolate from a patient in novosibirsk |
| title_short | Antigenic properties of sARs-CoV-2/human/RUs/nsk-FRCFtM-1/2020 coronavirus isolate from a patient in novosibirsk |
| title_sort | antigenic properties of sars cov 2 human rus nsk frcftm 1 2020 coronavirus isolate from a patient in novosibirsk |
| topic | коронавирус тяжелого острого респираторного синдрома 2 типа (sars-cov-2) коронавирусная инфекция 2019 (covid-19) культура клеток vero вирусные белки |
| url | https://journal.niidi.ru/jofin/article/view/1077 |
| work_keys_str_mv | AT aachepurnov antigenicpropertiesofsarscov2humanrusnskfrcftm12020coronavirusisolatefromapatientinnovosibirsk AT kasharshov antigenicpropertiesofsarscov2humanrusnskfrcftm12020coronavirusisolatefromapatientinnovosibirsk AT eikazachinskaya antigenicpropertiesofsarscov2humanrusnskfrcftm12020coronavirusisolatefromapatientinnovosibirsk AT yuvkononova antigenicpropertiesofsarscov2humanrusnskfrcftm12020coronavirusisolatefromapatientinnovosibirsk AT eakazachkova antigenicpropertiesofsarscov2humanrusnskfrcftm12020coronavirusisolatefromapatientinnovosibirsk AT opkhripko antigenicpropertiesofsarscov2humanrusnskfrcftm12020coronavirusisolatefromapatientinnovosibirsk AT ksyurchenko antigenicpropertiesofsarscov2humanrusnskfrcftm12020coronavirusisolatefromapatientinnovosibirsk AT ayualekseev antigenicpropertiesofsarscov2humanrusnskfrcftm12020coronavirusisolatefromapatientinnovosibirsk AT mivoevoda antigenicpropertiesofsarscov2humanrusnskfrcftm12020coronavirusisolatefromapatientinnovosibirsk AT amshestopalov antigenicpropertiesofsarscov2humanrusnskfrcftm12020coronavirusisolatefromapatientinnovosibirsk |