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

Woo Chang Chung, Kwang Yeon Hwang, Suk Jo Kang, Jae Ouk Kim, Moon Jung Song

Research output: Contribution to journalArticle

Abstract

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.

Original languageEnglish
Pages (from-to)46-53
Number of pages8
JournalJournal of Microbiology
Volume58
Issue number1
DOIs
Publication statusPublished - 2020 Jan 1

Fingerprint

Chikungunya virus
Polyproteins
Neutralizing Antibodies
Togaviridae
Alphavirus
Arboviruses
Neutralization Tests
Republic of Korea
Arthralgia
Virus Diseases
Exanthema
Culicidae
Luciferases
Reporter Genes
Human Activities
Antiviral Agents
Sequence Analysis
Headache
Fever
Vaccines

Keywords

  • Chikungunya virus
  • Human serum
  • Korean isolate
  • Neutralization assay
  • Pseudovirus

ASJC Scopus subject areas

  • Microbiology
  • Applied Microbiology and Biotechnology

Cite this

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.

In: Journal of Microbiology, Vol. 58, No. 1, 01.01.2020, p. 46-53.

Research output: Contribution to journalArticle

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