Coxsackievirus A10 (CV-A10) is one of the most common pathogens causing hand, foot and mouth disease (HFMD). The 39 candidate epitope peptides covering the amino acid sequence of the VP1 region of CV-A10 were screened by using the overlapping peptide method and verified by indirect enzyme linked immunosorbent assay (ELISA) and microneutral- ization inhibition assays. It has been found that candidate epitope P6 (amino acid 39~53 in VP1 region of CV-A10) showed high reactivity with CV-A10 virus serum, and still showed neutralizing inhibition effect at 3.91 μg/mL dilution mass concentration, which was a potential neutralizing epitope. P6 antiserum was prepared by immunizing mice with P6 peptide, and microneutral- ization test was used to verify its protective effect. The results showed a 1:8.97 geometric mean neutralizing titer for P6 antise- rum, which identified P6 as the neutralizing epitope of CV-A10. In order to understand the conservativeness of P6 sequence in CV-A10, the amino acid sequence of the P6 was compared with the representative strains of CV-A10 genotype, and the results showed that P6 was highly conserved in CV-A10. It indicated that P6 is a conserved linear neutralization epitope of CV-A10, which has the potential to resist the infection of various genotypes of CV-A10, and can be used as a target for the development of CV-A10 epitope vaccine. This study aimed to screen and identify linearly neutralizing epitopes on the CV-A10 VP1 protein and to lay a foundation for the development of CV-A10 epitope vaccine and HFMD prevention.