Guo Zhenyang, Zhang Siyu, Xu Hu, Li Wansheng, Li Chao, Zhao Jing, Gong Bangjun, Sun Qi, Xiang Lirun, Zhao Hongyuan, Wang Qian, Zhou Guohui, Tang Yandong, An Tongqing, Cai Xuehui, Tian Zhijun, Zhang Hongliang, Peng Jinmei
State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China.
School of Modern Agriculture and Biotechnology, Ankang University, Ankang 725000, China.
Vet Sci. 2023 Feb 9;10(2):133. doi: 10.3390/vetsci10020133.
Since 2011, pseudorabies virus (PRV) has recurred in several vaccinated pig farms in China. PRV variants with high virulence were found to be the main cause of the outbreaks. In the face of the PRV epidemic, detection of the wild strain is as important as vaccine immunization, so we hoped to achieve differential diagnosis of PRV by obtaining a monoclonal antibody (mAB) that could be used to identify the wild strain. In this study, we used a novel immunization and screening strategy to prepare an mAB and obtained mAB 1H5 against the gE glycoprotein. An immunofluorescence assay (IFA) revealed that this mAB was specific to both classic and variant strains of PRV. Subsequently, we further identified the linear epitopes of B cells recognized using the mAB. The mAB 1H5 bound at RRAG, which is a novel epitope and is conserved in almost all PRV strains. These findings provide novel insight into the structure and function of PRV proteins, the analysis of antigenic epitope characteristics, and the establishment of antigen or antibody detection methods.
自2011年以来,伪狂犬病病毒(PRV)在中国多个已接种疫苗的猪场中反复出现。高毒力的PRV变异株被发现是疫情爆发的主要原因。面对PRV疫情,野生毒株的检测与疫苗免疫同样重要,因此我们希望通过获得一种可用于鉴定野生毒株的单克隆抗体(mAB)来实现PRV的鉴别诊断。在本研究中,我们采用了一种新颖的免疫和筛选策略来制备单克隆抗体,并获得了针对gE糖蛋白的单克隆抗体1H5。免疫荧光分析(IFA)显示,该单克隆抗体对PRV的经典毒株和变异毒株均具有特异性。随后,我们进一步鉴定了使用该单克隆抗体识别的B细胞线性表位。单克隆抗体1H5结合于RRAG,这是一个新表位,几乎在所有PRV毒株中都保守。这些发现为PRV蛋白的结构和功能、抗原表位特征分析以及抗原或抗体检测方法的建立提供了新的见解。