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

Kelei Li, Fangyu Dong, Bopei Cui, Lisha Cui, Pei Liu, Chao Ma, Haifa Zheng, Xing Wu, Zhenglun Liang

Human vaccines & immunotherapeutics 2020 Jun 02

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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.

Citation

Kelei Li, Fangyu Dong, Bopei Cui, Lisha Cui, Pei Liu, Chao Ma, Haifa Zheng, Xing Wu, Zhenglun Liang. Development of a pseudovirus-based assay for measuring neutralizing antibodies against Coxsackievirus A10. Human vaccines & immunotherapeutics. 2020 Jun 02;16(6):1434-1440