Antigenic properties of sARs-CoV-2/human/RUs/nsk-FRCFtM-1/2020 coronavirus isolate from a patient in novosibirsk
| Autor: | K. S. Yurchenko, Yu V. Kononova, A. M. Shestopalov, K. A. Sharshov, M. I. Voevoda, A. Yu Alekseev, E. A. Kazachkova, O. P. Khripko, A. A. Chepurnov, E. I. Kazachinskaya |
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| Jazyk: | ruština |
| Rok vydání: | 2020 |
| Předmět: |
viruses
severe acute respiratory syndrome type 2 coronavirus (sars-cov-2) Infectious and parasitic diseases RC109-216 medicine.disease_cause 03 medical and health sciences Antigen medicine 030304 developmental biology Coronavirus Cytopathic effect chemistry.chemical_classification 0303 health sciences coronavirus infection 2019 (covid-19) Viral matrix protein biology 030306 microbiology Chemistry Virology Nucleoprotein Infectious Diseases viral proteins vero cell culture biology.protein Vero cell Antibody Glycoprotein |
| Zdroj: | Žurnal Infektologii, Vol 12, Iss 3, Pp 42-50 (2020) |
| ISSN: | 2072-6732 |
| Popis: | Objective:isolation of coronavirus SARS-CoV-2 fromclinical sample of patient with COVID-19 in Novosibirsk; obtaining a purified and inactivated viral antigen and study ofits antigenic properties. Materials and methods: virus isolation was carried out in Vero cell culture from nasopharyngealswab 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 RNAin cultural medium with reverse transcription – polymerasechain reaction (RT-PCR). Purification, concentration and inactivation of the viral preparation were carried out accordingto standard methods. The purity of the purified preparationand the profile of viral proteins were determined by electrophoresis in 10% polyacrylamide gel (PAG) with the additionof 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 antigenwas 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 inVero cell culture. The antigenic specificity of the three structural viral proteins (S, N, and M) is preserved in the purifiedinactivated viral preparation. The inactivated whole-virionantigen of SARS-CoV-2/human/RUS/Nsk-FRCFTM-1/2020isolate can be used to study the antigenic immunomodulating properties of viral proteins, to obtain immune sera oflaboratory animals, and also as a component of test systemsfor the detection of specific antibodies with ELISA and immunoblotting. |
| Databáze: | OpenAIRE |
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