TY - JOUR
T1 - A high-throughput yellow fever neutralization assay
AU - Rasulova, Madina
AU - Vercruysse, Thomas
AU - Paulissen, Jasmine
AU - Coun, Catherina
AU - Suin, Vanessa
AU - Heyndrickx, Leo
AU - Ma, Ji
AU - Geerts, Katrien
AU - Timmermans, Jolien
AU - Mishra, Niraj
AU - Li, Li-Hsin
AU - Kum, Dieudonné Buh
AU - Coelmont, Lotte
AU - Van Gucht, Steven
AU - Karimzadeh, Hadi
AU - Thorn-Seshold, Julia
AU - Rothenfußer, Simon
AU - Ariën, Kevin K
AU - Neyts, Johan
AU - Dallmeier, Kai
AU - Thibaut, Hendrik Jan
N1 - FTX; (CC BY 4.0)
PY - 2022
Y1 - 2022
N2 - 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.
AB - 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.
KW - Antibodies, Neutralizing
KW - Antibodies, Viral
KW - Encephalitis, Japanese
KW - Humans
KW - Neutralization Tests/methods
KW - Yellow Fever/diagnosis
KW - Yellow fever virus
KW - Zika Virus
KW - Zika Virus Infection
U2 - 10.1128/spectrum.02548-21
DO - 10.1128/spectrum.02548-21
M3 - A1: Web of Science-article
C2 - 35670599
SN - 2165-0497
VL - 10
JO - Microbiology Spectrum
JF - Microbiology Spectrum
IS - 3
M1 - e0254821
ER -