Development of a pseudovirus-based assay for measuring neutralizing antibodies against Coxsackievirus A10

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 timeconsuming 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.

Correlation between pseudotyped virus and authentic virus neutralisation assays, a systematic review and meta-analysis of the literature

“…Some of the studies used very small sample sizes, which was accounted for through giving lower weights to these studies. We opted to include studies that used PVs that are non-replicative, single cycle of infection, therefore excluding studies that used replicon infection systems, despite some of these reports showing high correlation and high level of agreement between single-round replicons and authentic virus in a neutralisation assay ( 110 , 111 ). Lastly, new virus and cell-free assays have now been developed for SARS-CoV-2 that measure the capability of antibodies blocking the spike protein from interacting with its receptor ACE-2, effectively becoming a surrogate neutralisation assay, have shown to have strong correlations with both pMNAs and vMNAs ( 51 , 69 , 112 – 114 ).…”

Identification of a neutralizing linear epitope within the VP1 protein of coxsackievirus A10

“…Coxsackievirus A10 (CV-A10) is a human enterovirus belonging to the A species of the Enterovirus genus within Picornaviridae family [ 1 ]. CV-A10-associated hand, foot, and mouth diseases (HFMD) typically manifest mild and self-limiting symptoms.…”