Practical aspects on identification, cultivation and characteristics of varicella-zoster virus isolates

Until now, it has been considered that infectivity of the varicella-zoster virus (VZV) is closely related to target cell, and newly formed virus is not released into the culture medium. It is also known that it is hard to grow VZV in cell cultures, due to its slow replication rate and a limited rang...

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Autores principales:	F. G. Nagieva, E. P. Barkova, A. N. Lisakov, A. V. Sidorov, V. V. Zverev, O. V. Osokina, A. D. Stroeva
Formato:	article
Lenguaje:	RU
Publicado:	Sankt-Peterburg : NIIÈM imeni Pastera 2020
Materias:	varicella-zoster virus cell culture сlinical isolate virus reproduction hemo-adsorbtion reaction immunofluorescent analysis monoclonal antibodies real-time pcr assay individual virus collecting Infectious and parasitic diseases RC109-216
Acceso en línea:	https://doaj.org/article/403ccf7cd73845c9bf5847665e5ed3bf
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spelling	oai:doaj.org-article:403ccf7cd73845c9bf5847665e5ed3bf2021-11-22T07:09:54ZPractical aspects on identification, cultivation and characteristics of varicella-zoster virus isolates2220-76192313-739810.15789/2220-7619-PAO-1211https://doaj.org/article/403ccf7cd73845c9bf5847665e5ed3bf2020-05-01T00:00:00Zhttps://www.iimmun.ru/iimm/article/view/1211https://doaj.org/toc/2220-7619https://doaj.org/toc/2313-7398Until now, it has been considered that infectivity of the varicella-zoster virus (VZV) is closely related to target cell, and newly formed virus is not released into the culture medium. It is also known that it is hard to grow VZV in cell cultures, due to its slow replication rate and a limited range of sensitive cell cultures. In addition, VZV isolation depends on type of cell culture used, nature of clinical material, presence of viable virus and transport time. Objectives. To study production of infectious extracellular VZV in various cell cultures. Materials and methods. Eight cell cultures were used, including human embryonic diploid lung cells and human embryonic dermomuscular tissue (KM-27), as well as continuous human and monkey cell lines. Crusts detached from vesicular lesions were used as clinical isolates, which were placed into cryo-vials added with transport medium and transferred in liquid nitrogen. VZV infectivity was assessed in cell cultures by using hemo-adsorption assay with erythrocyte suspension isolated from guinea pig or human zero group blood and confirmed by indirect immunofluorescence with polyclonal sera from varicella or herpes zoster convalescents. Results. There were examined 27 clinical samples consisting of crusts from vesicular lesions isolated from patients with chickenpox, as well as one sample from 63-year old patient with exacerbated recurrent herpes zoster. Primary infection with clinical isolates was performed on diploid human lung embryo cells (HLEC) at low temperature. It was found that clinical samples collected within day 1–18 inclusive after the onset of skin eruption were able to induce cytopathic effects in HLEC cell monolayer such as cytolysis around dermal crusts. Specificity of cytopathic effect was confirmed by using real-time polymerase chain reaction (RT-PCR). Viral antigens were prepared on 7 cell lines infected with the laboratory strain Ellen VZV (USA) to assess the immune sera. A high anti-VZV specificity of mouse sera was detected by ELISA while all the lysates of infected cell lines were used as the solid-phase sorbent. In experiments on VZV reproduction demonstrated that extracellular virus was released into the culture medium starting from day 1 after infection of target cells, and infectivity of the virus-containing fluid ascends during further cultivation.F. G. NagievaE. P. BarkovaA. N. LisakovA. V. SidorovV. V. ZverevO. V. OsokinaA. D. StroevaSankt-Peterburg : NIIÈM imeni Pasteraarticlevaricella-zoster viruscell cultureсlinical isolatevirus reproductionhemo-adsorbtion reactionimmunofluorescent analysismonoclonal antibodiesreal-time pcr assayindividual virus collectingInfectious and parasitic diseasesRC109-216RUInfekciâ i Immunitet, Vol 10, Iss 2, Pp 387-396 (2020)
institution	DOAJ
collection	DOAJ
language	RU
topic	varicella-zoster virus cell culture сlinical isolate virus reproduction hemo-adsorbtion reaction immunofluorescent analysis monoclonal antibodies real-time pcr assay individual virus collecting Infectious and parasitic diseases RC109-216
spellingShingle	varicella-zoster virus cell culture сlinical isolate virus reproduction hemo-adsorbtion reaction immunofluorescent analysis monoclonal antibodies real-time pcr assay individual virus collecting Infectious and parasitic diseases RC109-216 F. G. Nagieva E. P. Barkova A. N. Lisakov A. V. Sidorov V. V. Zverev O. V. Osokina A. D. Stroeva Practical aspects on identification, cultivation and characteristics of varicella-zoster virus isolates
description	Until now, it has been considered that infectivity of the varicella-zoster virus (VZV) is closely related to target cell, and newly formed virus is not released into the culture medium. It is also known that it is hard to grow VZV in cell cultures, due to its slow replication rate and a limited range of sensitive cell cultures. In addition, VZV isolation depends on type of cell culture used, nature of clinical material, presence of viable virus and transport time. Objectives. To study production of infectious extracellular VZV in various cell cultures. Materials and methods. Eight cell cultures were used, including human embryonic diploid lung cells and human embryonic dermomuscular tissue (KM-27), as well as continuous human and monkey cell lines. Crusts detached from vesicular lesions were used as clinical isolates, which were placed into cryo-vials added with transport medium and transferred in liquid nitrogen. VZV infectivity was assessed in cell cultures by using hemo-adsorption assay with erythrocyte suspension isolated from guinea pig or human zero group blood and confirmed by indirect immunofluorescence with polyclonal sera from varicella or herpes zoster convalescents. Results. There were examined 27 clinical samples consisting of crusts from vesicular lesions isolated from patients with chickenpox, as well as one sample from 63-year old patient with exacerbated recurrent herpes zoster. Primary infection with clinical isolates was performed on diploid human lung embryo cells (HLEC) at low temperature. It was found that clinical samples collected within day 1–18 inclusive after the onset of skin eruption were able to induce cytopathic effects in HLEC cell monolayer such as cytolysis around dermal crusts. Specificity of cytopathic effect was confirmed by using real-time polymerase chain reaction (RT-PCR). Viral antigens were prepared on 7 cell lines infected with the laboratory strain Ellen VZV (USA) to assess the immune sera. A high anti-VZV specificity of mouse sera was detected by ELISA while all the lysates of infected cell lines were used as the solid-phase sorbent. In experiments on VZV reproduction demonstrated that extracellular virus was released into the culture medium starting from day 1 after infection of target cells, and infectivity of the virus-containing fluid ascends during further cultivation.
format	article
author	F. G. Nagieva E. P. Barkova A. N. Lisakov A. V. Sidorov V. V. Zverev O. V. Osokina A. D. Stroeva
author_facet	F. G. Nagieva E. P. Barkova A. N. Lisakov A. V. Sidorov V. V. Zverev O. V. Osokina A. D. Stroeva
author_sort	F. G. Nagieva
title	Practical aspects on identification, cultivation and characteristics of varicella-zoster virus isolates
title_short	Practical aspects on identification, cultivation and characteristics of varicella-zoster virus isolates
title_full	Practical aspects on identification, cultivation and characteristics of varicella-zoster virus isolates
title_fullStr	Practical aspects on identification, cultivation and characteristics of varicella-zoster virus isolates
title_full_unstemmed	Practical aspects on identification, cultivation and characteristics of varicella-zoster virus isolates
title_sort	practical aspects on identification, cultivation and characteristics of varicella-zoster virus isolates
publisher	Sankt-Peterburg : NIIÈM imeni Pastera
publishDate	2020
url	https://doaj.org/article/403ccf7cd73845c9bf5847665e5ed3bf
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Practical aspects on identification, cultivation and characteristics of varicella-zoster virus isolates

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