Zhang Zhaohan, Gu Wenyuan, Fu Xiaotian, Lin Jiayu, Ding Xiuyan, Zhu Liqian
Engineering Research Center of Microbial Breeding and Preservation of Hebei Province, College of Life Sciences, Hebei University, Baoding, 071002, China.
Center for Animal Diseases Control and Prevention of Hebei Province, Shijiazhuang, 050035, China.
BMC Microbiol. 2025 Feb 10;25(1):70. doi: 10.1186/s12866-025-03782-2.
BoAHV1 is a significant cattle pathogen, resulting in substantial economic losses worldwide. B-cell epitopes within the viral proteins are not well understood. In this study, we screened a 12-mer phage display peptide library using a commercial BoAHV1 antibody. A total of 16 phage clones displaying individual 12-mer peptides were identified as reactive with this antibody using dot bot analysis. Through sequence alignment, 10 putative BoAHV1 B-cell epitopes, designated as PV109, PV108, PV116, PV59, PV50, PV130, PV113, PV49, PV62, and PV133, located within viral proteins gB, gC, gG, gM, UL36, UL37, and UL49, respectively, were recognized by both commercial BoAHV1 antibody andBoAHV1 IgG positive cattle serum through dot blot assay. Interestingly, immunization of mice with the synthesized peptide PV116 led to the production of antibodies suitable for Western blot analysis. Furthermore, an ELISA kit for the detection of BoAHV1 IgG antibodies in serum was developed, utilizing the identified epitope PV49, PV108, PV109, and PV116 as coating antigen. Collectively, we have identified 10 novel B-cell epitopes ofBoAHV1. Among them, PV116 is capable of inducing the production of antibodies suitable for Western blotting assay, which also shows potential for the development diagnostic tools.
牛α疱疹病毒1型(BoAHV1)是一种重要的牛病原体,在全球范围内造成了巨大的经济损失。病毒蛋白中的B细胞表位尚未得到充分了解。在本研究中,我们使用市售的BoAHV1抗体筛选了一个12肽噬菌体展示肽库。通过斑点印迹分析,共鉴定出16个展示单个12肽的噬菌体克隆与该抗体发生反应。通过序列比对,位于病毒蛋白gB、gC、gG、gM、UL36、UL37和UL49内的10个假定的BoAHV1 B细胞表位,分别命名为PV109、PV108、PV116、PV59、PV50、PV130、PV113、PV49、PV62和PV133,通过斑点印迹法被市售的BoAHV1抗体和BoAHV1 IgG阳性牛血清所识别。有趣的是,用合成肽PV116免疫小鼠可产生适用于蛋白质印迹分析的抗体。此外,利用鉴定出的表位PV49、PV108、PV109和PV116作为包被抗原,开发了一种用于检测血清中BoAHV1 IgG抗体的ELISA试剂盒。我们总共鉴定出了10个新的BoAHV1 B细胞表位。其中,PV116能够诱导产生适用于蛋白质印迹分析的抗体,这也显示出其在开发诊断工具方面的潜力。