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HE Yunyi¹, LlU Yangyang^2a , HU Jiahua^2b,3, LIU Hongbo^1,2a

( 1.College of Medical Laboratory Science, Guilin Medical University, Guilin 541199, China; 2. a. Department ofLaboratory Medicine , b. Department of Science and Research, the Second AffiliatedHospital of Guilin Medical University, Guilin 541199, China; 3.Guangdong Provincial Center forDisease Control and Prevention,Cuangzhou 5l1430.China)

Abstract Objective 'To identify linear B cell epitopes of VPi protein of enterovirus C96(EV-C96). Methods Thelinear B cell epitopes and the property parameters of EV-C96 VPl protein were predicted by the lEDB onlineanalysis tools, combined with its secondary structure parameters such as B-uming angle, antigenicity, andhydrophilicity were used to sereen out candidate epitopes with advantages. The reactivity of the candidate epitopeswith the EV-C96 antiserum and the cross-reactivity with antiserums against 2l other common enterovirus caused handfoot and mouth disease were determined by indirect ELlSA assay. The neutralization titer of epitope antibody wasdetermined by microneutralization test. The conservativeness of epitopes was analyzed by sequence alignment , andthe structural specificity of epitopes was analyzed by structural alignment. Results Seven epitopes of EV-C96 VP1protein( P 1 ~ P7) were predicted by bioinformatics methods, among which P7( aa282 ~ 304) showed strong reactivitywith EV-C96 antiserum and did not cross-react with other common enterovirus antisera. The neutralizing titer of P7antilbody was less than 1 : 2. The amino acid sequence of P7 was highly conservative in EV-C96 isolates, and thestructure of the P7 was diflerent from other 21 common EVs. Conclusion P7 is a non-neutral EV-C96-specific B cellepitope that can be used as a candidate antigen epitope for the development of EV-C96 detection kits.Keywords: enterovirus C96; VPl protein ; linear B cell epitopes; hand-foot-and-mouth disease

DOI:10.19296/j.cnki.1008-2409.2024-02-001

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