Abstract
Diseases caused by the genus Flavivirus, including dengue virus (DENV) and Zika virus (ZIKV), have a serious impact on public health worldwide. Due to serological cross-reactivity among flaviviruses, current enzyme-linked immunosorbent assay (ELISA) for IgM/G cannot reliably distinguish between infection by different flaviviruses. In this study, we developed a reporter-based neutralization assay using single-round infectious particles (SRIPs) derived from representative flaviviruses. SRIPs were generated by transfection of human embryonic kidney 293 T cells with a plasmid encoding premembrane and envelope (prME) proteins from DENV1–4, ZIKV, Japanese encephalitis virus, West Nile virus, yellow fever virus, Usutu virus, and tick-borne encephalitis virus, along with a plasmid carrying DENV1 replicon containing the luciferase gene and plasmid for expression of DENV1 capsid. Luciferase activity of SRIPs-infected cells was well correlated with number of infected cells, and each reporter SRIP was specifically neutralized by sera from mice immunized with each flavivirus antigen. Our high-throughput reporter SRIP-based neutralization assay for multiple flaviviruses is a faster, safer, and less laborious diagnostic method than the conventional plaque reduction neutralization test to screen the cause of primary flavivirus infection. The assay may also contribute to the evaluation of vaccine efficacy and assist in routine surveillance and outbreak response to flaviviruses.
Highlights
Many flaviviruses cause significant human morbidity and mortality, including dengue virus (DENV), Zika virus (ZIKV), yellow fever virus (YFV), West Nile virus (WNV), Japanese encephalitis virus (JEV), and tick-borne encephalitis virus (TBEV), and are transmitted by mosquitoes or ticks
We previously reported that single-round infectious particles (SRIPs) for JEV, DENV, and ZIKV can be successfully used in place of authentic viruses in neutralization tests with an advantage in terms of safety[12,13,14] because SRIP-infected cells are not able to produce progeny virions due to the lack of partial structural gene in the packaged genome
We demonstrate the utility of multiple flavivirus reporter SRIPs as an alternative tool to authentic flaviviruses in neutralization assays
Summary
Many flaviviruses cause significant human morbidity and mortality, including dengue virus (DENV), Zika virus (ZIKV), yellow fever virus (YFV), West Nile virus (WNV), Japanese encephalitis virus (JEV), and tick-borne encephalitis virus (TBEV), and are transmitted by mosquitoes or ticks. A single open reading frame encodes the viral polyprotein, which is processed into structural [capsid (C), premembrane/membrane (prM/M), and envelope (E)] and nonstructural (NS1, NS2A, NS2B, NS3, NS4A, NS4B, and NS5) proteins by viral and host proteases. This open reading frame is flanked by 5′ and 3′ untranslated regions. We previously reported that single-round infectious particles (SRIPs) for JEV, DENV, and ZIKV can be successfully used in place of authentic viruses in neutralization tests with an advantage in terms of safety[12,13,14] because SRIP-infected cells are not able to produce progeny virions due to the lack of partial structural gene in the packaged genome. We demonstrate that reporter SRIPs from several flaviviruses can be used as viral particles for neutralization assays as a faster and safe diagnostic method of flavivirus infection
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
