microbeПроблемы особо опасных инфекцийProblems of Particularly Dangerous Infections0370-10692658-719XRussian Research Anti-Plague Institute “Microbe”10.21055/0370-1069-2022-1-94-100microbe-1662Research ArticleОРИГИНАЛЬНЫЕ СТАТЬИORIGINAL ARTICLESИзучение в условиях in vitro биологических свойств штаммов коронавируса SARS-CoV-2, относящихся к различным генетическим вариантамIn vitro Study of Biological Properties of SARS-CoV-2 Coronavirus Strains Related to Various Genetic Variantshttps://orcid.org/0000-0002-0450-5212ЗайковскаяА. В.ZaykovskayaA. V.

Зайковская Анна Владимировна

630559, Новосибирская обл., р.п. Кольцово

Kol’tsovo, Novosibirsk Region, 630559

zaykovskaya_av@vector.nsc.ruhttps://orcid.org/0000-0002-7396-3954ГладышеваА. В.GladyshevaA. V.

630559, Новосибирская обл., р.п. Кольцово

Kol’tsovo, Novosibirsk Region, 630559

https://orcid.org/0000-0002-7857-6822КарташовМ. Ю.KartashovM. Yu.

630559, Новосибирская обл., р.п. Кольцово

Kol’tsovo, Novosibirsk Region, 630559

https://orcid.org/0000-0002-6746-8092ТарановО. С.TaranovO. S.

630559, Новосибирская обл., р.п. Кольцово

Kol’tsovo, Novosibirsk Region, 630559

https://orcid.org/0000-0002-1745-7643ОвчинниковаА. С.OvchinnikovaA. S.

630559, Новосибирская обл., р.п. Кольцово

Kol’tsovo, Novosibirsk Region, 630559

https://orcid.org/0000-0003-1201-8307ШиповаловА. В.ShipovalovА. V.

630559, Новосибирская обл., р.п. Кольцово

Kol’tsovo, Novosibirsk Region, 630559

https://orcid.org/0000-0003-3340-8750ПьянковО. В.P’yankovO. V.

630559, Новосибирская обл., р.п. Кольцово

Kol’tsovo, Novosibirsk Region, 630559

ФБУН Государственный научный центр вирусологии и биотехнологии ВекторРоссияState Scientific Center of Virology and Biotechnology VectorRussian Federation2022190420220194100Copyright © Зайковская А.В., Гладышева А.В., Карташов М.Ю., Таранов О.С., Овчинникова А.С., Шиповалов А.В., Пьянков О.В., 20222022Зайковская А.В., Гладышева А.В., Карташов М.Ю., Таранов О.С., Овчинникова А.С., Шиповалов А.В., Пьянков О.В.Zaykovskaya A.V., Gladysheva A.V., Kartashov M.Y., Taranov O.S., Ovchinnikova A.S., Shipovalov А.V., P’yankov O.V.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://journal.microbe.ru/jour/article/view/1662

Цель – изучить особенности репродукции штаммов вируса SARS-CoV-2 различных генетических вариантов на культурах клеток Vero и Vero E6.

Материалы и методы

Материалы и методы. В работе использовали штаммы вируса SARS-CoV-2, относящиеся к вариантам, вызывающим обеспокоенность (VOC), циркулирующим на территории РФ. Штаммы вируса SARS-CoV-2 депонированы в Государственной коллекции возбудителей вирусных инфекций и риккетсиозов ФБУН ГНЦ ВБ «Вектор» Роспотребнадзора. Эксперименты проводили на культурах клеток Vero и Vero E6. Динамику накопления инфекционного вируса определяли путем титрования образцов культуральной жидкости через 24, 48, 72, 96 часов после инфицирования (MOI – от 1 до 0,00001 ЦПД50/клетку). Образование бляшек изучали на культуре клеток Vero E6 под 0,2 % агаровым покрытием. Анализ изображения и подсчет размеров бляшек проводили в графическом редакторе GIMP (GNU Image Manipulation Program).

Результаты и обсуждение

Результаты и обсуждение. Описана динамика накопления инфекционного вируса в культуральной жидкости в зависимости от множественности инфицирования для штаммов вируса SARS-CoV-2, относящихся к разным генетическим линиям. Показаны различия морфологии бляшек на монослое культуры клеток Vero E6 под агаровым покрытием. Вирусы SARS-CoV-2, относящиеся к генетическим вариантам альфа и дельта, демонстрируют максимальную репродукцию среди изученных штаммов (инфекционный титр – более 7 lg ТЦД50/100мкл). Вариант омикрон под агаровым покрытием образует самые мелкие бляшки и при низкой множественности инфицирования имеет низкий уровень репродукции. Таким образом, штаммы коронавируса SARS-CoV-2, относящиеся к разным генетическим линиям, имеют существенные отличия в скорости репродукции на культуре клеток Vero и Vero Е6.

The aim of the research was to study reproduction features of SARS-CoV-2 coronavirus strains of various genetic lines on Vero and Vero E6 cell culture.

Materials and methods

Materials and methods. The SARS-CoV-2 virus strains related to the variants of concern (VOC) circulating in the territory of the Russian Federation were used in the research. The strains of the SARS-CoV-2 virus were deposited in the State Collection of Pathogens of Viral Infections and Rickettsioses at the FBIS SSC VB “Vector” of the Rospotrebnadzor. The experiments were carried out on Vero and Vero E6 cell cultures. The dynamics of infectious virus accumulation was determined by titration of culture fluid samples 24, 48, 72, 96 hours after infection (MOI – from 1 to 0,00001 CPE50/cell). Plaque formation was studied on Vero E6 cell culture under 0.2 % agar coating. Image analysis and plaque size calculation were performed using GIMP (GNU Image Manipulation Program).

Results and discussion

Results and discussion. The study describes the dynamics of accumulation of infectious virus in the culture fluid depending upon multiplicity of infection for the strains of SARS-CoV-2 virus belonging to different genetic lines. Differences in the morphology of plaques on the monolayer of Vero E6 cell culture under agar coating are shown. SARS-CoV-2 virus strains related to Alfa and Delta VOC demonstrate maximum reproduction rate among the studied strains (infectious titer is higher than 7 lg TCID50/100µl). Omicron VOC forms small plaques under agar coating and at a low multiplicity of infection has a low reproduction rate. Thus, SARS-CoV-2 virus strains belonging to different genetic lines have significant differences in the rate of reproduction on Vero and Vero E6 cell culture.

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