The development and validation of a microneutralization assay for the detection and quantification of anti-yellow fever virus antibodies in human serum.

Unlabelled: The plaque reduction neutralization test (PRNT) has been used widely for the detection and quantitation of yellow fever (YF) virus-neutralizing antibodies in human serum; however, it is labor-intensive and challenging to adapt to high-throughput clinical testing needed for vaccine licensure. Here, we describe the development and validation of a new Vero cell-based YF microneutralization (MN) assay, with immunostaining readout, for the detection and quantification of YF virus-neutralizing antibodies in human serum. Comparison of neutralizing antibody titers measured with the YF MN assay versus the historical YF PRNT, based on a 50% reduction in plaque counts (PRNT), demonstrated 100% serostatus agreement at a titer of 10 (1/dil) in participants with a history of YF vaccination.