Development of a Coxsackievirus A16 neutralization assay based on pseudoviruses for measurement of neutralizing antibody titer in human serum

► Non-proliferating CV-A16 pseudoviruses were generated by reverse genetics. ► A novel pseudovirus-based CV-A16 neutralization assay (pCNA) was developed. ► The kinetics of CVA16 pseudovirus infectivity was determined in vitro. ► Screening with the pCNA correlated well with a conventional neutralization assay. ► The pCNA may be a rapid screening test for CV-A16 infection.