He Kuijing, Wu Yue, Su Zhipeng, Zeng Yue, Ye Guishan, Wu Qi, Li Long, Zhang Anding
State Key Laboratory of Agricultural Microbiology, Hubei Hongshan Laboratory, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China.
Key Laboratory of Preventive Veterinary Medicine in Hubei Province, The Cooperative Innovation Center for Sustainable Pig Production, Wuhan 430070, China.
Viruses. 2025 Jun 7;17(6):823. doi: 10.3390/v17060823.
African swine fever virus (ASFV) is a highly virulent pathogen that causes nearly 100% mortality in acute infections and poses persistent risks. Effective containment of ASFV outbreaks requires rapid and reliable diagnostic tools. The p54 protein, a key structural component of ASFV, has emerged as an important target for serological detection. Herein, the recombinant p54 protein (amino acids 53-184) was expressed in , and three mouse monoclonal antibodies (mAbs) (IgG1/kappa subtype) were developed. Among these mAbs, the mAb 1F9 specifically recognized the B-cell epitope IQFINPYQDQQ, which is conserved across different genotypes of ASFV, suggesting that the epitope may serve as a valuable target for serological detection of ASFV. Structural modeling analysis revealed that this epitope is surface-exposed on the p54 protein, with Gln and Phe identified as critical residues for 1F9 binding. Moreover, a blocking ELISA based on the mAb 1F9 was established for detecting ASFV-specific antibodies in clinical serum samples, achieving a coincidence rate exceeding 95%. These findings demonstrate that mAb 1F9, targeting a conserved and accessible region of p54, represents a valuable tool for ASFV serodiagnosis, surveillance, and outbreak management.
非洲猪瘟病毒(ASFV)是一种高致病性病原体,在急性感染中可导致近100%的死亡率,并带来持续风险。有效控制ASFV疫情需要快速且可靠的诊断工具。p54蛋白是ASFV的关键结构成分,已成为血清学检测的重要靶点。在此,重组p54蛋白(氨基酸53 - 184)在[具体表达系统未给出]中表达,并制备了三种小鼠单克隆抗体(mAb)(IgG1/kappa亚型)。在这些单克隆抗体中,mAb 1F9特异性识别B细胞表位IQFINPYQDQQ,该表位在不同基因型的ASFV中保守,这表明该表位可能是ASFV血清学检测的重要靶点。结构建模分析表明,该表位在p54蛋白表面暴露,Gln和Phe被确定为1F9结合的关键残基。此外,基于mAb 1F9建立了阻断ELISA法用于检测临床血清样本中的ASFV特异性抗体,符合率超过95%。这些结果表明,靶向p54保守且易接近区域的mAb 1F9是ASFV血清学诊断、监测和疫情管理的重要工具。