（1. 桂林医学院第二附属医院检验科，桂林 541199；2. 桂林医学院医学检验学院，桂林 541199；3. 桂林医学院智能医学与生物技术学院，桂林 541199）

摘　要：柯萨奇病毒A组10型（CV-A10）是引起手足口病（HFMD）的常见病原体之一。应用重叠肽法筛选出覆盖CV-A10全长 VP1区氨基酸序列的39条候选表位肽段，以间接酶联免疫吸附试验（ELISA）及微量中和抑制试验验证。结果发现，候选表位P6（CV-A10 VP1区第39～53位氨基酸）与CV-A10全病毒抗血清具有高反应性，且在3.91 μg/mL的稀释质量浓度时仍具有中和抑制作用，为潜在中和表位。用P6肽免疫小鼠制备相应的抗血清，并以微量中和试验验证其保护能力，结果显示，P6抗血清的几何平均中和效价为1:8.97，可确定P6为CV-A10的中和表位。为了解P6序列在CV-A10型内的保守性，将P6的氨基酸序列与CV-A10各基因型代表株进行比对分析，结果显示，P6在CV-A10型内高度保守，表明P6为CV-A10的广谱性中和表位，可应用于CV-A10表位疫苗的研发。本研究旨在筛选鉴定CV-A10 VP1蛋白上的线性中和表位，为CV-A10表位疫苗的研发和HFMD的防控奠定基础。

关键词：柯萨奇病毒A组10型；VP1蛋白；线性中和表位；疫苗；手足口病

中图分类号：R373.2；R512.5                    文献标志码：A DOI：10.3969/j.issn.1007-7146.2024.02.008

(1. Department of Laboratory Medicine, the Second Affiliated Hospital of Guilin Medical University, Guilin 541199, China; 2. College of Medical Laboratory Science, Guilin Medical University, Guilin 541199, China; 3. College of Intelligent Medicine and Biotechnology, Guilin Medical University, Guilin 541199, China)

Abstract: 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 microneutralization 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 microneutralization test was used to verify its protective effect. The results showed a 1:8.97 geometric mean neutralizing titer for P6 antiserum, 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.

Key words: coxsackievirus A10; VP1 protein; linear neutral epitope; vaccine; hand, foot and mouth disease

（Acta Laser Biology Sinica, 2024, 33(2): 160-166）