Xia Nengwen, Cao Qi, Liu Anjing, Zhang Jiajia, Han Hongjian, Jiao Jun, He Ping, Sun Ziyan, Xu Zijian, Zheng Wanglong, Jiang Sen, Chen Nanhua, Bai Jianfa, Zhu Jianzhong
College Veterinary Medicine, Yangzhou University, Yangzhou 225009, China.
Joint International Research Laboratory of Agriculture and Agri-Product Safety, Yangzhou University, Yangzhou 225009, China.
Vet Sci. 2024 Dec 13;11(12):650. doi: 10.3390/vetsci11120650.
African swine fever (ASF) has widely spread around the world in the last 100 years since its discovery. The African swine fever virus (ASFV) particles are made of more than 150 proteins, with the p17 protein encoded by the D117L gene serving as one of the major capsid proteins and playing a crucial role in the virus's morphogenesis and immune evasion. Thus, monoclonal antibody (mAb) targeting p17 is important for the research and detection of ASFV infection. Here, we produced two specific mAbs against p17, designated as 1G2 and 6G3, respectively, and both have been successfully used in enzyme-linked immunosorbent assay (ELISA), Western blotting, and immunofluorescence assay. Moreover, we found that both 1G2 and 6G3 mAbs recognize a novel epitope of 72-78 amino acids of p17 protein, highly conserved across all genotype I and II strains. Based on this epitope, an indirect ELISA has been established to effectively detect antibodies during ASFV infection, and it exhibits high consistency with commercial ASFV ELISA kits. In summary, the production of the specific p17 mAbs and the identification of the recognized epitope will significantly promote the serological diagnosis of ASFV.
自发现以来的近100年里，非洲猪瘟（ASF）已在全球广泛传播。非洲猪瘟病毒（ASFV）颗粒由150多种蛋白质组成，其中由D117L基因编码的p17蛋白是主要衣壳蛋白之一，在病毒的形态发生和免疫逃避中起关键作用。因此，靶向p17的单克隆抗体（mAb）对ASFV感染的研究和检测具有重要意义。在此，我们制备了两种针对p17的特异性单克隆抗体，分别命名为1G2和6G3，二者均已成功应用于酶联免疫吸附测定（ELISA）、蛋白质免疫印迹法和免疫荧光测定。此外，我们发现1G2和6G3单克隆抗体均识别p17蛋白72 - 78位氨基酸的一个新表位，该表位在所有I型和II型基因型毒株中高度保守。基于此表位，已建立一种间接ELISA方法以有效检测ASFV感染期间的抗体，并且它与市售ASFV ELISA试剂盒具有高度一致性。总之，特异性p17单克隆抗体的产生以及所识别表位的鉴定将显著促进ASFV的血清学诊断。
