A high-throughput yellow fever neutralization assay

Madina Rasulova, Thomas Vercruysse, Jasmine Paulissen, Catherina Coun, Vanessa Suin, Leo Heyndrickx, Ji Ma, Katrien Geerts, Jolien Timmermans, Niraj Mishra, Li-Hsin Li, Dieudonné Buh Kum, Lotte Coelmont, Steven Van Gucht, Hadi Karimzadeh, Julia Thorn-Seshold, Simon Rothenfußer, Kevin K Ariën, Johan Neyts, Kai DallmeierHendrik Jan Thibaut

Research output: Contribution to journalA1: Web of Science-article

Abstract

Quick and accurate detection of neutralizing antibodies (nAbs) against yellow fever is essential in serodiagnosis during outbreaks for surveillance and to evaluate vaccine efficacy in population-wide studies. All of this requires serological assays that can process a large number of samples in a highly standardized format. Albeit being laborious, time-consuming, and limited in throughput, the classical plaque reduction neutralization test (PRNT) is still considered the gold standard for the detection and quantification of nAbs due to its sensitivity and specificity. Here, we report the development of an alternative fluorescence-based serological assay (SNTFLUO) with an equally high sensitivity and specificity that is fit for high-throughput testing with the potential for automation. Finally, our novel SNTFLUO was cross-validated in several reference laboratories and against international WHO standards, showing its potential to be implemented in clinical use. SNTFLUO assays with similar performance are available for the Japanese encephalitis, Zika, and dengue viruses amenable to differential diagnostics. IMPORTANCE Fast and accurate detection of neutralizing antibodies (nAbs) against yellow fever virus (YFV) is key in yellow fever serodiagnosis, outbreak surveillance, and monitoring of vaccine efficacy. Although classical PRNT remains the gold standard for measuring YFV nAbs, this methodology suffers from inherent limitations such as low throughput and overall high labor intensity. We present a novel fluorescence-based serum neutralization test (SNTFLUO) with equally high sensitivity and specificity that is fit for processing a large number of samples in a highly standardized manner and has the potential to be implemented for clinical use. In addition, we present SNTFLUO assays with similar performance for Japanese encephalitis, Zika, and dengue viruses, opening new avenues for differential diagnostics.

Original languageEnglish
Article numbere0254821
JournalMicrobiology Spectrum
Number of pages11
ISSN2165-0497
DOIs
Publication statusE-pub ahead of print - 2022

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