Zhang Jianhua, Huang Huiqi, Xu Lian, Lou Chaonan, Pan Mi
Department of Laboratory Medicine, School of Medicine, Shaoxing University, Shaoxing, P.R. China.
Viral Immunol. 2019 Mar;32(2):84-88. doi: 10.1089/vim.2018.0125. Epub 2018 Dec 6.
Enterovirus 71 (EV-A71) has attracted widespread attention in the past decade because of its higher neurotropicity in addition to causing hand-foot-and-mouth disease (HFMD). Mapping epitopes of viral proteins may promote our understanding of antiviral humoral immunity, and is useful for clinical application. In this study, the linear B cell epitopes within nonstructural proteins of EV-A71 were identified using bioinformatics methods combined with peptide enzyme-linked immunosorbent assay (ELISA). Twenty epitopes were predicted and located at 2ABC (7), 3ABC (5), and 3D (8) protein, respectively. Of 20 epitope-containing peptides, 14 were verified by ELISA (S/CO >2.1), 9 of which had higher reactivity (S/CO >5.0). Furthermore, synthetic peptide SP09 (EV-A71-3ABC) can react with healthy children sera, and its immunoreactivity was closest to that of EV-A71-VP1 protein. The protein BLAST analysis revealed that SP09 may contain a common epitope due to the high homology of amino acid sequences with other members of human Enterovirus species A. These findings may be useful in the development serological tests for the diagnosis of HFMD caused by a broad range of human Enterovirus species A.
肠道病毒71型（EV - A71）在过去十年中引起了广泛关注，因为它除了引发手足口病（HFMD）外，还具有更高的嗜神经性。绘制病毒蛋白的表位可能有助于我们理解抗病毒体液免疫，并且对临床应用有用。在本研究中，结合肽酶联免疫吸附测定（ELISA）使用生物信息学方法鉴定了EV - A71非结构蛋白内的线性B细胞表位。预测了20个表位，分别位于2ABC（7个）、3ABC（5个）和3D（8个）蛋白上。在20个含表位的肽中，14个通过ELISA得到验证（S/CO>2.1），其中9个具有更高的反应性（S/CO>5.0）。此外，合成肽SP09（EV - A71 - 3ABC）可与健康儿童血清发生反应，其免疫反应性最接近EV - A71 - VP1蛋白。蛋白质BLAST分析显示，由于氨基酸序列与人类肠道病毒A种的其他成员具有高度同源性，SP09可能包含一个共同表位。这些发现可能有助于开发用于诊断由多种人类肠道病毒A种引起的手足口病的血清学检测方法。
