ArticlePLoS neglected tropical diseases2025
Assessing anti-rabies vaccine response in humans: A rapid and high-throughput adaptable, pseudovirus-based neutralization assay as an alternative to rapid fluorescent focus inhibition test (RFFIT).
Article in PLoS neglected tropical diseases, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 2 papers.
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Who cites it
2 citing papers in PubMed.
- Enhanced immunogenicity of the rabies virus glycoprotein fused to a dendritic cell-targeting peptide.Scientific reports · 2026Article
- Reproducible generation of Nipah virus pseudovirions with uniform incorporation of F and G surface glycoproteins for high-throughput neutralization assays.BMC microbiology · 2025Article
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Authors and funding
7 authors.
Funding
Abstract
backgroundRabies is a fatal, but vaccine-preventable disease. The conventional 'gold standard' recommended for estimating anti-rabies vaccine immune response is Rapid Fluorescent Focus Inhibition Test (RFFIT). Here, we describe a pseudovirus-based neutralization assay (PVNT) as an alternative method to detect the neutralizing antibody response. METHODOLOGY/PRINCIPAL
findingsWe used a recombinant Vesicular Stomatitis Virus (rVSV)-based pseudotyping system in HEK293 cells transfected with rabies G glycoprotein to produce high titre pseudoviruses. These were confirmed by neutralization with anti-G monoclonal antibodies and a reference serum, as well as immunofluorescence-based detection of pseudovirus infection. A secreted alkaline phosphatase (SEAP) reporter was included to make the assay high-throughput adaptable with readouts possible using a conventional ELISA reader. A PVNT was developed that had a dynamic range of OD405 from 0.10 to 2.00, and was used to test a set of RFFIT-validated 71 human serum samples. Results of PVNT and RFFIT showed significant concordance with a Pearson's correlation value of 0.8555. In samples with border-line RFFIT positive values (a RFFIT titre in the range of 0.5-1.8 IU/mL), concordance was low, with the PVNT results tending to be negative. PVNT assay had a specificity of 100% when compared to RFFIT. The assay also had 100% positive predicative value indicating its robustness for use in the evaluation for the presence of neutralizing antibodies against a near 100% fatal disease such as rabies. CONCLUSIONS/SIGNIFICANCE: Our results show that PVNT is a reproducible, sensitive and specific assay with an objective and documentable readout. The assay is faster with a turnaround time of less than 24h and can be automated for large scale screening. Importantly, it can be performed without the restrictions in handling live rabies virus. PVNT provides a reliable alternative to RFFIT for monitoring vaccine efficacy and immunity status, making it a valuable tool in rabies control programs.
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