ArticleMethodsX2026
NanoLuc-based microneutralization assay for high-throughput detection of neutralizing antibodies against rift valley fever virus.
Article in MethodsX, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
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3 authors.
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Abstract
Plaque reduction neutralization tests (PRNTs) are the gold standard for measuring neutralizing antibodies but are labor-intensive, low-throughput, and subject to variability from manual plaque counting. We developed a NanoLuc-based microneutralization (mNT) assay using rMP12-NanoLuc that quantifies luminescence in culture supernatants as a surrogate for infection. The assay runs in 96-well plates without overlay, fixation, or staining, using 35 focus-forming unit (FFU) per well and a 40 hour readout with back-titration to ensure accuracy. mNT titers from vaccinated animal sera showed strong correlation with conventional PRNTs. This approach enables rapid, reproducible, high-throughput detection of RVFV neutralizing antibodies, with the main limitations being substrate cost and the requirement for a luminometer.•The assay enables high-throughput, objective neutralization testing in 96-well plates.•Optimized conditions yield mNT titers that strongly correlate with conventional PRNTs.•Limitations include NanoLuc substrate cost and the need for a luminometer.
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