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

用于功能性抗体检测的荧光素酶分泌、单轮传染性 DENV-2 报告基因的产生和表征

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作者: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

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.

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