Cao Yimei, Li Fengjuan, Xing Xiangchuan, Zhang Huiyan, Zhao QiongQiong, Sun Pu, Fu Yuanfang, Li Pinghua, Ma Xueqing, Zhang Jing, Zhao Zhixun, Yuan Hong, Wang Jian, Wang Tao, Bao Huifang, Bai Xingwen, Li Dong, Zhang Qiang, Li Kun, Lu Zengjun
State Key Laboratory for Animal Disease Control and Prevention, College of Veterinary Medicine, Lanzhou University, National Foot-and-Mouth Diseases Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, China.
Microbiol Spectr. 2025 Feb 4;13(2):e0223424. doi: 10.1128/spectrum.02234-24. Epub 2025 Jan 8.
Neutralizing antibodies provide vital protection against foot-and-mouth disease virus (FMDV). The virus neutralization test (VNT) is a gold standard method for the detection of neutralizing antibodies. However, its application is limited due to the requirement for live virus and unsuitability for large-scale serological surveillance. In this study, a porcine broadly neutralizing monoclonal antibody (PO18-10) against FMDV was obtained from the heterologous sequentially vaccinated pig using single-B-cell antibody technology. A competitive enzyme-linked immunosorbent assay (C-ELISA) for detecting neutralizing antibodies against FMDV serotype O was developed using biotinylated PO18-10 as a detector antibody. The sensitivity and specificity of the assay were 100% and 99.55%, respectively, and the positive/negative coincidence rate with VNT was 94%, suggesting that C-ELISA based on natural host-derived monoclonal antibody (mAb) could be a promising tool to detect neutralizing antibodies against FMDV serotype O and evaluate the vaccine efficacy.IMPORTANCEFoot-and-mouth disease virus (FMDV) serotype O is one of the most prevalent serotypes in the world. The neutralizing antibody titers in primo-vaccinated animals are directly related to their level of protection against a virus challenge. The development of a safe, rapid, and accurate method for the detection of the neutralizing antibody is essential for the control and eradication of FMD. In this study, an inter-serotype broadly neutralizing monoclonal antibody PO18-10 was successfully produced using single-B-cell antibody technology from sequentially vaccinated pigs. A competitive ELISA based on this natural host-derived mAb for the detection of neutralizing antibodies against FMDV serotype O was developed and validated. The assay demonstrates high sensitivity, specificity, and coincidence rate with VNT, making it an alternative tool for confirming FMDV infection and evaluating the vaccine efficacy.
中和抗体为抵御口蹄疫病毒（FMDV）提供至关重要的保护。病毒中和试验（VNT）是检测中和抗体的金标准方法。然而，由于需要活病毒且不适用于大规模血清学监测，其应用受到限制。在本研究中，利用单B细胞抗体技术从经异源顺序接种的猪中获得了一种针对FMDV的猪源广谱中和单克隆抗体（PO18 - 10）。以生物素化的PO18 - 10作为检测抗体，开发了一种用于检测抗口蹄疫病毒O型中和抗体的竞争酶联免疫吸附测定（C - ELISA）。该测定的灵敏度和特异性分别为100%和99.55%，与VNT的阳性/阴性符合率为94%，这表明基于天然宿主来源单克隆抗体（mAb）的C - ELISA可能是检测抗口蹄疫病毒O型中和抗体及评估疫苗效力的一种有前景的工具。
重要性
口蹄疫病毒（FMDV）O型是世界上最流行的血清型之一。初次接种疫苗动物体内的中和抗体滴度与其抵御病毒攻击的保护水平直接相关。开发一种安全、快速且准确的检测中和抗体的方法对于口蹄疫的控制和根除至关重要。在本研究中，利用单B细胞抗体技术从顺序接种疫苗的猪中成功制备了一种跨血清型广谱中和单克隆抗体PO18 - 10。开发并验证了一种基于这种天然宿主来源mAb的竞争ELISA，用于检测抗口蹄疫病毒O型中和抗体。该测定显示出高灵敏度、特异性以及与VNT的符合率，使其成为确认FMDV感染和评估疫苗效力的替代工具。
