Abstract
The lack of reliable, high-throughput tools for characterizing anti-dengue virus (DENV) antibodies in large numbers of serum samples has been an obstacle in understanding the impact of neutralizing antibodies on disease progression and vaccine efficacy. A reporter system using pseudoinfectious DENV reporter virus particles (RVPs) was previously developed by others to facilitate the genetic manipulation and biological characterization of DENV virions. In the current study, we demonstrate the diagnostic utility of DENV RVPs for measuring neutralizing antibodies in human serum samples against all four DENV serotypes, with attention to the suitability of DENV RVPs for large-scale, long-term studies. DENV RVPs used against human sera yielded serotype-specific responses and reproducible neutralization titers that were in statistical agreement with Plaque Reduction Neutralization Test (PRNT) results. DENV RVPs were also used to measure neutralization titers against the four DENV serotypes in a panel of human sera from a clinical study of dengue patients. The high-throughput capability, stability, rapidity, and reproducibility of assays using DENV RVPs offer advantages for detecting immune responses that can be applied to large-scale clinical studies of DENV infection and vaccination.
Highlights
Dengue virus (DENV) is a member of the Flavivirus genus in the family Flaviviridae and consists of four distinct serotypes (DENV-1, DENV-2, DENV-3, and DENV-4), which are transmitted by Ae. aegypti and Ae. albopictus mosquitoes
We developed DENV reporter virus particles (RVPs) against all 4 dengue serotypes that are antigenically equivalent to commonlyused strains in DENV research and vaccine design: DENV-1 (WestPac), DENV-2 (S16803), DENV-3 (CH53489), and DENV-4 (TVP360)
We demonstrate the production of DENV RVPs that are antigenically equivalent to each of the four DENV serotypes, facilitating the detection of neutralizing activity in human sera
Summary
Dengue virus (DENV) is a member of the Flavivirus genus in the family Flaviviridae and consists of four distinct serotypes (DENV-1, DENV-2, DENV-3, and DENV-4), which are transmitted by Ae. aegypti and Ae. albopictus mosquitoes. Primary DENV infection may confer lifelong protection from re-infection with the same serotype, it only provides shortterm protection from infection with additional serotypes [4], highlighting the need for a dengue vaccine that can confer persistent and simultaneous protection against all four serotypes of the virus. The development of such a vaccine continues to be a significant challenge, made more difficult by the lack of efficient and reliable screening methods for detecting and evaluating functional antibodies in human sera
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