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Author Spotlight: Studying Host-Virus Interactions with Pseudotyped Viruses
Published on: November 21, 2023
Development of a pseudovirus-based assay for measuring neutralizing antibodies against Coxsackievirus A10
Kelei Li1,2, Fangyu Dong1,3, Bopei Cui1
1Division of Hepatitis Virus Vaccines, National Institute for Food and Drug Control , Beijing, China.
Insights
A new pseudovirus neutralization assay (pNT) offers a rapid and efficient method for detecting neutralizing antibodies against Coxsackievirus A10 (CV-A10). This assay significantly speeds up vaccine development for hand, foot, and mouth disease.
Area of Science:
- Virology
- Immunology
- Vaccine Development
Background:
- Coxsackievirus A10 (CV-A10) is a significant cause of hand, foot, and mouth disease in children globally.
- Effective treatments for CV-A10 infections are lacking, making vaccine development a priority.
- Accurate measurement of neutralizing antibodies (NtAb) is crucial for advancing CV-A10 vaccine research.
Purpose of the Study:
- To develop an efficient, high-throughput assay for measuring neutralizing antibodies against CV-A10.
- To compare the performance of the new pseudovirus-based neutralization assay (pNT) with the traditional cytopathic effect inhibition assay (cNT).
Main Methods:
- Development of a CV-A10 pseudovirus (pNT) assay utilizing a luciferase reporter gene.
- Quantification of anti-CV-A10 NtAb titres based on relative luminescent units (RLU).
- Parallel testing of 67 human serum samples using both pNT and cNT assays.
Main Results:
- The pNT assay detected anti-CV-A10 NtAb within 10-16 hours.
- A strong positive correlation (r=0.83, p<0.0001) and high agreement (97%) were observed between the pNT and cNT assays.
- The pNT assay demonstrated a rapid and convenient procedure for NtAb measurement.
Conclusions:
- The pseudovirus-based neutralization assay (pNT) is a validated, efficient, and high-throughput method for measuring neutralizing antibodies against CV-A10.
- This assay will accelerate the development of effective CV-A10 vaccines.
- The pNT assay provides a reliable alternative to traditional, labor-intensive neutralization assays.
Abstract:
Coxsackievirus A10 (CV-A10) has recently emerged as a major pathogen of hand, foot, and mouth disease in children worldwide. Currently no effective treatments are available; development of anti-CV-A10 vaccine is a most cost-effective way for CV-A10 prevention. Robust assay to measure neutralizing antibody (NtAb) titres elicited by vaccination would greatly prompt anti-CV-A10 vaccine development. Compare to the traditional neutralization assay based on inhibition of cytopathic effects (herein after referred to as cNT) which is time-consuming and labor-intensive, in this study we developed an efficient high-throughput neutralization antibody assay based on CV-A10 pseudoviruses (herein after referred to as pNT). In the pNT, anti-CV-A10 NtAb titre was negatively corresponded with the relative luminescent unit (RLU) produced by luciferase reporter gene incorporated in pseudovirus genome. As described in this study, the NtAb against CV-A10 could be detected within 10-16 h, anti- CV-A10 NtAb in 67 human serum samples were measured in parallel with pNT and cNT assays, a good correlation (r = 0.83,p < .0001) and good agreement(97%) were shown between cNT and pNT, indicating that the pNT provides a rapid and convenient procedure for measuring NtAb production against anti-CV-A10 NtAb measurement.
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