Generation and characterization of luciferase-secreting, single-round infectious DENV-2 reporter for functional antibody assays

Jiraphan Junjhon; Kedsara Panyasu; Suwipa Chaiyaloom; Krongkan Saipin; Pornsiri Somasa; Sutha Sangiambut; Chunya Puttikhunt; Rungtawan Sriburi; Poonsook Keelapang; Peeraya Ekchariyawat; Panisadee Avirutnan; Chakrit Hirunpetcharat; Nopporn Sittisombut

doi:10.1016/j.jviromet.2021.114119

Generation and characterization of luciferase-secreting, single-round infectious DENV-2 reporter for functional antibody assays

J Virol Methods. 2021 May:291:114119. doi: 10.1016/j.jviromet.2021.114119. Epub 2021 Mar 1.

Affiliations

¹ Department of Microbiology, Faculty of Public Health, Mahidol University, Bangkok, 10400, Thailand. Electronic address: jiraphan.juh@mahidol.ac.th.
² Department of Microbiology, Faculty of Public Health, Mahidol University, Bangkok, 10400, Thailand.
³ Molecular Biology of Dengue and Flaviviruses Research Team, Medical Molecular Biotechnology Research Group, National Center for Genetic Engineering and Biotechnology, National Science and Technology Development Agency, Pathum-thani, 12120, Thailand; Division of Dengue Hemorrhagic Fever Research and Siriraj Center of Research Excellence in Dengue and Emerging Pathogens, Faculty of Medicine, Siriraj Hospital, Mahidol University, Bangkok, 10700, Thailand.
⁴ Department of Microbiology, Faculty of Medicine, Chiang Mai University, Chiang Mai, 50200, Thailand.
⁵ Division of Dengue Hemorrhagic Fever Research and Siriraj Center of Research Excellence in Dengue and Emerging Pathogens, Faculty of Medicine, Siriraj Hospital, Mahidol University, Bangkok, 10700, Thailand.
⁶ Molecular Biology of Dengue and Flaviviruses Research Team, Medical Molecular Biotechnology Research Group, National Center for Genetic Engineering and Biotechnology, National Science and Technology Development Agency, Pathum-thani, 12120, Thailand; Department of Microbiology, Faculty of Medicine, Chiang Mai University, Chiang Mai, 50200, Thailand.

PMID: 33662412
DOI: 10.1016/j.jviromet.2021.114119

Abstract

Flavivirus reporters provide a robust tool for viral pathogenesis studies, anti-viral drug screening, disease diagnosis and functional antibody assays. In this study, we generated a luciferase-secreting, single-round reporter virus by replacing the capsid coding region in a DENV-2 genome with the secretory form of Lucia luciferase gene to produce infectious viral particles in a stable capsid-expressing mosquito cell line. Replication of the reporter virus in trans-complementing mosquito cells was sustained for up to two weeks. There were strong correlations between the extracellular luciferase activity and infectious reporter virus inocula upon infection of mosquito and mammalian cell lines with graded quantities of the reporter virus. A set of anti-E and anti-prM monoclonal antibodies affected the infectivity of reporter virus with similar dose-effect relationships as the parent virus. This simplified version of DENV-2 reporter provides a rapid and reliable method for the detection of neutralizing and infection-enhancing antibodies against dengue virus.

Keywords: Antibody assay; Dengue virus; Reporter virus; Secreted luciferase.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Antibodies, Blocking
Antibodies, Viral
Dengue Virus* / genetics
Dengue* / diagnosis
Flavivirus*
Luciferases / genetics

Substances

Antibodies, Blocking
Antibodies, Viral
Luciferases