A High-Throughput Yellow Fever Neutralization Assay

Abstract

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.

Live-attenuated YF17D-vectored COVID-19 vaccine protects from lethal yellow fever virus infection in mouse and hamster models

“… 18 , 28 [preprint] YFV nAb were determined using mCherry-tagged YF17D reporter viruses. 18 , 28 [preprint, 29 ] All reagents were prepared and methods were performed accordingly.…”

Validation of a Reporter Cell Line for Flavivirus Inhibition Assays

“…4 ). The sera used were obtained from patients suspected of having ZIKV infection or from nonhuman primates (NHPs) vaccinated against YFV that had previously been assessed by the Arbovirus National Reference Center at the Institute of Tropical Medicine (ITM) using a PRNT ( 13 ). Viruses were incubated with serum dilutions before addition to the reporter cell line, luciferase activity was measured after 72 h, and inhibition profiles were fitted with a four-parameter variable-slope curve.…”

“…Monkey sera were collected from rhesus macaques vaccinated with the licensed YF17D vaccine Stamaril ( 13 ).…”

Multiplexed multicolor antiviral assay amenable for high-throughput research

“…All viruses used were generated recombinantly from infectious cDNA clones. The construction and characterization of mCherry-tagged YF17D (YF17D/mCherry) and eGFP-tagged JEV SA14-14-2 strains (JEV/eGFP) have been reported previously 60 – 62 . The construction of infectious clone pShuttle-DV2/mCherry expressing the full-length RNA genome of DENV-2 New Guinea C (NGC) strain plus mCherry as N-terminal translational fusion to the DENV-2 polyprotein has been described earlier 23 .…”