A novel reporter system for neutralizing and enhancing antibody assay against dengue virus
- Equal contributors
1 Department of Virology, State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing 100071, China
2 Department of Microbiology and Immunology, School of Medicine Jinan University, Guangzhou 510632, China
3 Novartis Institute for Tropical Diseases, Singapore 138670, Singapore
4 State Key Laboratory of Virology, Wuhan Institute of Virology, CAS, Wuhan 430072, China
5 Guangzhou No.8 People’s Hospital, Guangzhou 510060, China
BMC Microbiology 2014, 14:44 doi:10.1186/1471-2180-14-44Published: 18 February 2014
Dengue virus (DENV) still poses a global public health threat, and no vaccine or antiviral therapy is currently available. Antibody plays distinct roles in controlling DENV infections. Neutralizing antibody is protective against DENV infection, whereas sub-neutralizing concentration of antibody can increase DENV infection, termed antibody-dependent enhancement (ADE). Plaque-based assay represents the most widely accepted method measuring neutralizing or enhancing antibodies.
In this study, a novel reporter virus-based system was developed for measuring neutralization and ADE activity. A stable Renilla luciferase reporter DENV (Luc-DENV) that can produce robust luciferase signals in BHK-21 and K562 cells were used to establish the assay and validated against traditional plaque-based assay. Luciferase value analysis using various known DENV-specific monoclonal antibodies showed good repeatability and a well linear correlation with conventional plaque-based assays. The newly developed assay was finally validated with clinical samples from infected animals and individuals.
This reporter virus-based assay for neutralizing and enhancing antibody evaluation is rapid, lower cost, and high throughput, and will be helpful for laboratory detection and epidemiological investigation for DENV antibodies.