Utility of an In-Vitro Micro-Neutralizing Test in Comparison to a Plaque Reduction Neutralization Test for Dengue Virus, Japanese Encephalitis Virus, and Zika Virus Serology and Drug Screening

• Type of Material: Article
• Authors: Kazumi Haga, University of Tokyo; Zhenying (Nancy) Chen, Emory University; Misao Himeno, University of Tokyo; Ryuichi Majima, University of Tokyo; Meng Ling Moi, University of Tokyo
• Language: English
• Date: 2023-12-20
• Publisher: MDPI
• Publication Version: Final Published Version
• Copyright Statement: © 2023 by the authors.
• License: Creative Commons Attribution 4.0 International
• Final Published Version (URL): https://doi.org/10.3390/pathogens13010008
• Title of Journal or Parent Work: Pathogens
• Volume: 13
• Issue: 1
• Start Page: 8
• Grant/Funding Agency: Japan Agency for Medical Research and Development
• Grant/Funding Information: This research was supported by the Japan Agency for Medical Research and Development (AMED) under grants JP233fa627001, JP22fk0108123 and 23wm0225033h0001.
• Keywords: dengue; neutralizing antibodies; flavivirus; microneutralization test; Japanese encephalitis virus; Zika
• Subject - Topics: Virology; World health

Abstract

Flavivirus infections, including dengue virus (DENV), Japanese encephalitis virus (JEV), and Zika virus (ZIKV), present significant global public health challenges. For successful vaccine design, the assessment of neutralizing antibody activity requires reliable and robust methodologies for determining antibody titers. Although the plaque reduction neutralization test (PRNT) is commonly acknowledged as the gold standard, it has limitations in terms of time and cost, and its usage may be limited in resource-limited settings. To address these challenges, we introduced the micro-neutralization test (MNT) as a simplified alternative to the PRNT. The MNT employs a 96-well plate format, conducts microscale neutralization assays, and assesses cell viability by dissolving cells to create a uniform color solution, which is measured with a spectrometer. In this study, we evaluated the utility of the MNT by contrasting the end-point titers of the MNT and PRNT using 4 monoclonal antibodies, 15 non-human primate serum samples, and 2 therapeutic drug candidates across flaviviruses. The results demonstrated a strong correlation between the MNT and PRNT titers, affirming the robustness and reproducibility of the MNT for evaluating control measures against flaviviruses. This research contributes valuable insights toward the development of a cost-effective antibody titer testing approach that is particularly suitable for resource-limited settings.