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

• Published in: Journal of Microbiology Vol. 58; no. 1; pp. 46 - 53
• Main Authors: Chung, Woo-Chang, Hwang, Kwang Yeon, Kang, Suk-Jo, Kim, Jae-Ouk, Song, Moon Jung
• Format: Journal Article
• Language: English
• Published: Seoul Springer Science and Business Media LLC 01.01.2020; The Microbiological Society of Korea; Springer Nature B.V; 한국미생물학회
• Subjects: Antibodies; Antibodies, Neutralizing; Antibodies, Neutralizing - blood; Antibodies, Viral; Antibodies, Viral - blood; Antiviral agents; Antiviral drugs; Aquatic insects; arboviruses; Arthralgia; Assaying; Biomedical and Life Sciences; Biosafety; Chikungunya Fever; Chikungunya Fever - immunology; Chikungunya Fever - virology; Chikungunya virus; Chikungunya virus - immunology; Chikungunya virus - isolation & purification; disease outbreaks; Epidemics; Fever; Fluorescence; Genes; Genomes; genotype; Genotypes; Global health; green fluorescent protein; Headache; Humans; Life Sciences; luciferase; Microbiological strains; Microbiology; Neutralization; Neutralization Tests; Neutralization Tests - methods; Neutralizing; neutralizing antibodies; Nucleic acids; Pathogens; Phylogeny; polyproteins; rapid methods; reporter genes; Republic of Korea; Ribonucleic acid; RNA; Sequencing; South Korea; Test procedures; Vaccines; Vector-borne diseases; Virology; Viruses; 생물학; Republic of Korea; South Korea; Human serum; Neutralization assay; Korean isolate; Chikungunya virus; Pseudovirus
• ISSN: 1225-8873; 1976-3794; 1976-3794
• DOI: 10.1007/s12275-020-9384-0

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.