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Development of a neutralization assay based on the pseudotyped chikungunya virus of a Korean isolate

The Chikungunya virus (CHIKV) belongs to the Alphavirus genus of Togaviridae family and contains a positive-sense single stranded RNA genome. Infection by this virus mainly causes sudden high fever, rashes, headache, and severe joint pain that can last for several months or years. CHIKV, a mosquito-...

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Autores principales: Chung, Woo-Chang, Hwang, Kwang Yeon, Kang, Suk-Jo, Kim, Jae-Ouk, Song, Moon Jung
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Microbiological Society of Korea 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7091072/
https://www.ncbi.nlm.nih.gov/pubmed/31768937
http://dx.doi.org/10.1007/s12275-020-9384-0
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author Chung, Woo-Chang
Hwang, Kwang Yeon
Kang, Suk-Jo
Kim, Jae-Ouk
Song, Moon Jung
author_facet Chung, Woo-Chang
Hwang, Kwang Yeon
Kang, Suk-Jo
Kim, Jae-Ouk
Song, Moon Jung
author_sort Chung, Woo-Chang
collection PubMed
description The Chikungunya virus (CHIKV) belongs to the Alphavirus genus of Togaviridae family and contains a positive-sense single stranded RNA genome. Infection by this virus mainly causes sudden high fever, rashes, headache, and severe joint pain that can last for several months or years. CHIKV, a mosquito-borne arbovirus, is considered a re-emerging pathogen that has become one of the most pressing global health concerns due to a rapid increase in epidemics. Because handling of CHIKV is restricted to Biosafety Level 3 (BSL-3) facilities, the evaluation of prophylactic vaccines or antivirals has been substantially hampered. In this study, we first iden-tified the whole structural polyprotein sequence of a CHIKV strain isolated in South Korea (KNIH/2009/77). Phylogenetic analysis showed that this sequence clustered within the East/ Central/South African CHIKV genotype. Using this sequence information, we constructed a CHIKV-pseudotyped lenti-virus expressing the structural polyprotein of the Korean CHIKV isolate (CHIKVpseudo) and dual reporter genes of green fluorescence protein and luciferase. We then developed a pseudovirus-based neutralization assay (PBNA) using CHIKVpseudo. Results from this assay compared to those from the conventional plaque reduction neutralization test showed that our PBNA was a reliable and rapid method to evaluate the efficacy of neutralizing antibodies. More importantly, the neutralizing activities of human sera from CHIKV-infected individuals were quantitated by PBNA using CHIKVpseudo. Taken together, these results suggest that our PBNA for CHIKV may serve as a useful and safe method for testing the neutralizing activity of antibodies against CHIKV in BSL-2 facilities.
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spelling pubmed-70910722020-03-24 Development of a neutralization assay based on the pseudotyped chikungunya virus of a Korean isolate Chung, Woo-Chang Hwang, Kwang Yeon Kang, Suk-Jo Kim, Jae-Ouk Song, Moon Jung J Microbiol Virology The Chikungunya virus (CHIKV) belongs to the Alphavirus genus of Togaviridae family and contains a positive-sense single stranded RNA genome. Infection by this virus mainly causes sudden high fever, rashes, headache, and severe joint pain that can last for several months or years. CHIKV, a mosquito-borne arbovirus, is considered a re-emerging pathogen that has become one of the most pressing global health concerns due to a rapid increase in epidemics. Because handling of CHIKV is restricted to Biosafety Level 3 (BSL-3) facilities, the evaluation of prophylactic vaccines or antivirals has been substantially hampered. In this study, we first iden-tified the whole structural polyprotein sequence of a CHIKV strain isolated in South Korea (KNIH/2009/77). Phylogenetic analysis showed that this sequence clustered within the East/ Central/South African CHIKV genotype. Using this sequence information, we constructed a CHIKV-pseudotyped lenti-virus expressing the structural polyprotein of the Korean CHIKV isolate (CHIKVpseudo) and dual reporter genes of green fluorescence protein and luciferase. We then developed a pseudovirus-based neutralization assay (PBNA) using CHIKVpseudo. Results from this assay compared to those from the conventional plaque reduction neutralization test showed that our PBNA was a reliable and rapid method to evaluate the efficacy of neutralizing antibodies. More importantly, the neutralizing activities of human sera from CHIKV-infected individuals were quantitated by PBNA using CHIKVpseudo. Taken together, these results suggest that our PBNA for CHIKV may serve as a useful and safe method for testing the neutralizing activity of antibodies against CHIKV in BSL-2 facilities. The Microbiological Society of Korea 2019-11-25 2020 /pmc/articles/PMC7091072/ /pubmed/31768937 http://dx.doi.org/10.1007/s12275-020-9384-0 Text en © The Microbiological Society of Korea 2020 This article is made available via the PMC Open Access Subset for unrestricted research re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic.
spellingShingle Virology
Chung, Woo-Chang
Hwang, Kwang Yeon
Kang, Suk-Jo
Kim, Jae-Ouk
Song, Moon Jung
Development of a neutralization assay based on the pseudotyped chikungunya virus of a Korean isolate
title Development of a neutralization assay based on the pseudotyped chikungunya virus of a Korean isolate
title_full Development of a neutralization assay based on the pseudotyped chikungunya virus of a Korean isolate
title_fullStr Development of a neutralization assay based on the pseudotyped chikungunya virus of a Korean isolate
title_full_unstemmed Development of a neutralization assay based on the pseudotyped chikungunya virus of a Korean isolate
title_short Development of a neutralization assay based on the pseudotyped chikungunya virus of a Korean isolate
title_sort development of a neutralization assay based on the pseudotyped chikungunya virus of a korean isolate
topic Virology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7091072/
https://www.ncbi.nlm.nih.gov/pubmed/31768937
http://dx.doi.org/10.1007/s12275-020-9384-0
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