Virological and genetic characteristics of human respiratory syncytial viruses isolated in Russia, 2017-2018

Isolation of human respiratory syncytial virus (HRSV) from clinical samples and storage of isolates for long period remains a considerable problem. We describe in detail the optimized conditions of HRSV isolation and cultivation in three cell cultures HeLa, HEp-2, and Vero. HRSV was detected in 35....

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Main Authors: Elena Alexandrovna Prokopyeva, Olga Grigoryevna Kurskaya, Mariya Vladimirovna Solomatina, Ivan Andreevich Sobolev, Tereza Araevna Saroyan, Nikita Artemyevich Dubovitskiy, Anastasiya Alexandrovna Derko, Peter Mikchailovich Kozhin, Tatyana Yurievna Alikina, Marsel Rasimovich Kabilov, Alexander Mikchailovich Shestopalov, Kirill Alexandrovich Sharshov
Format: Article
Language:English
Published: The Journal of Infection in Developing Countries 2023-02-01
Series:Journal of Infection in Developing Countries
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Online Access:https://jidc.org/index.php/journal/article/view/17462
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author Elena Alexandrovna Prokopyeva
Olga Grigoryevna Kurskaya
Mariya Vladimirovna Solomatina
Ivan Andreevich Sobolev
Tereza Araevna Saroyan
Nikita Artemyevich Dubovitskiy
Anastasiya Alexandrovna Derko
Peter Mikchailovich Kozhin
Tatyana Yurievna Alikina
Marsel Rasimovich Kabilov
Alexander Mikchailovich Shestopalov
Kirill Alexandrovich Sharshov
author_facet Elena Alexandrovna Prokopyeva
Olga Grigoryevna Kurskaya
Mariya Vladimirovna Solomatina
Ivan Andreevich Sobolev
Tereza Araevna Saroyan
Nikita Artemyevich Dubovitskiy
Anastasiya Alexandrovna Derko
Peter Mikchailovich Kozhin
Tatyana Yurievna Alikina
Marsel Rasimovich Kabilov
Alexander Mikchailovich Shestopalov
Kirill Alexandrovich Sharshov
author_sort Elena Alexandrovna Prokopyeva
collection DOAJ
description Isolation of human respiratory syncytial virus (HRSV) from clinical samples and storage of isolates for long period remains a considerable problem. We describe in detail the optimized conditions of HRSV isolation and cultivation in three cell cultures HeLa, HEp-2, and Vero. HRSV was detected in 35.2% (166/471) specimens by real-time PCR from symptomatic infants and children up to 15 years from October 2017 to March 2018 in Russia. HRSV-positive samples were used for virus isolation in HeLa, HEp-2, and Vero cells in different manners (in monolayer or suspension). To optimize the conditions of HRSV cultivation, these cell cultures were treated or not with receptor-destroying enzyme (RDE). Ten isolates were successfully obtained by the way of infection of the suspension of cells with subsequent RDE treatment. Among them, several isolates induced the cytopathogenic effect (CPE) by the syncytium formation in both Hela and HEp-2 cell cultures. The genetic analysis revealed that the manners of isolation by using monolayer or suspension and subsequent RDE treatment did not influence the nucleotide and amino acid structures of obtained HRSVs. The CPE characteristics of obtained viruses were the same in HeLa, HEp-2, and Vero cell cultures, and were described as large syncytium up to 150 microns or more in size with the nuclei peripheral location and an optically bright zone in the center of the formation. We showed that infection of cell suspension with the subsequent RDE treatment increased the chance of HRSVs isolation from clinical samples.
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spelling doaj-art-56d85bee4fb74a39ba80aa9f88edf6982025-08-20T03:48:47ZengThe Journal of Infection in Developing CountriesJournal of Infection in Developing Countries1972-26802023-02-01170210.3855/jidc.17462Virological and genetic characteristics of human respiratory syncytial viruses isolated in Russia, 2017-2018Elena Alexandrovna Prokopyeva0Olga Grigoryevna Kurskaya1Mariya Vladimirovna Solomatina2Ivan Andreevich Sobolev3Tereza Araevna Saroyan4Nikita Artemyevich Dubovitskiy5Anastasiya Alexandrovna Derko6Peter Mikchailovich Kozhin7Tatyana Yurievna Alikina8Marsel Rasimovich Kabilov9Alexander Mikchailovich Shestopalov10Kirill Alexandrovich Sharshov11Federal Research Center of Fundamental and Translational Medicine, Novosibirsk, RussiaFederal Research Center of Fundamental and Translational Medicine, Novosibirsk, RussiaFederal Research Center of Fundamental and Translational Medicine, Novosibirsk, RussiaFederal Research Center of Fundamental and Translational Medicine, Novosibirsk, RussiaFederal Research Center of Fundamental and Translational Medicine, Novosibirsk, RussiaFederal Research Center of Fundamental and Translational Medicine, Novosibirsk, RussiaFederal Research Center of Fundamental and Translational Medicine, Novosibirsk, RussiaFederal Research Center of Fundamental and Translational Medicine, Novosibirsk, RussiaInstitute of Chemical Biology and Fundamental Medicine, Novosibirsk, RussiaInstitute of Chemical Biology and Fundamental Medicine, Novosibirsk, RussiaFederal Research Center of Fundamental and Translational Medicine, Novosibirsk, RussiaFederal Research Center of Fundamental and Translational Medicine, Novosibirsk, Russia Isolation of human respiratory syncytial virus (HRSV) from clinical samples and storage of isolates for long period remains a considerable problem. We describe in detail the optimized conditions of HRSV isolation and cultivation in three cell cultures HeLa, HEp-2, and Vero. HRSV was detected in 35.2% (166/471) specimens by real-time PCR from symptomatic infants and children up to 15 years from October 2017 to March 2018 in Russia. HRSV-positive samples were used for virus isolation in HeLa, HEp-2, and Vero cells in different manners (in monolayer or suspension). To optimize the conditions of HRSV cultivation, these cell cultures were treated or not with receptor-destroying enzyme (RDE). Ten isolates were successfully obtained by the way of infection of the suspension of cells with subsequent RDE treatment. Among them, several isolates induced the cytopathogenic effect (CPE) by the syncytium formation in both Hela and HEp-2 cell cultures. The genetic analysis revealed that the manners of isolation by using monolayer or suspension and subsequent RDE treatment did not influence the nucleotide and amino acid structures of obtained HRSVs. The CPE characteristics of obtained viruses were the same in HeLa, HEp-2, and Vero cell cultures, and were described as large syncytium up to 150 microns or more in size with the nuclei peripheral location and an optically bright zone in the center of the formation. We showed that infection of cell suspension with the subsequent RDE treatment increased the chance of HRSVs isolation from clinical samples. https://jidc.org/index.php/journal/article/view/17462Human respiratory syncytial virus infectioncultivationisolationsuspensionmonolayersequences
spellingShingle Elena Alexandrovna Prokopyeva
Olga Grigoryevna Kurskaya
Mariya Vladimirovna Solomatina
Ivan Andreevich Sobolev
Tereza Araevna Saroyan
Nikita Artemyevich Dubovitskiy
Anastasiya Alexandrovna Derko
Peter Mikchailovich Kozhin
Tatyana Yurievna Alikina
Marsel Rasimovich Kabilov
Alexander Mikchailovich Shestopalov
Kirill Alexandrovich Sharshov
Virological and genetic characteristics of human respiratory syncytial viruses isolated in Russia, 2017-2018
Journal of Infection in Developing Countries
Human respiratory syncytial virus infection
cultivation
isolation
suspension
monolayer
sequences
title Virological and genetic characteristics of human respiratory syncytial viruses isolated in Russia, 2017-2018
title_full Virological and genetic characteristics of human respiratory syncytial viruses isolated in Russia, 2017-2018
title_fullStr Virological and genetic characteristics of human respiratory syncytial viruses isolated in Russia, 2017-2018
title_full_unstemmed Virological and genetic characteristics of human respiratory syncytial viruses isolated in Russia, 2017-2018
title_short Virological and genetic characteristics of human respiratory syncytial viruses isolated in Russia, 2017-2018
title_sort virological and genetic characteristics of human respiratory syncytial viruses isolated in russia 2017 2018
topic Human respiratory syncytial virus infection
cultivation
isolation
suspension
monolayer
sequences
url https://jidc.org/index.php/journal/article/view/17462
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