Wang Junli, Lei Baishi, Zhang Wuchao, Li Lijie, Ji Jiashuang, Liu Mandi, Zhao Kuan, Yuan Wanzhe
College of Veterinary Medicine, Hebei Agricultural University, Baoding 071000, China.
Hebei Veterinary Biotechnology Innovation Center, Hebei Agricultural University, Baoding 071000, China.
Animals (Basel). 2024 Jan 11;14(2):235. doi: 10.3390/ani14020235.
Porcine circovirus type 3 (PCV3) is endemic in swine worldwide and causes reproductive disorders, dermatitis and nephrotic syndrome, and multi-organ inflammation. Currently, there is a growing need for rapid and accurate diagnostic methods in disease monitoring. In this study, four monoclonal antibodies (mAbs) against PCV3 capsid proteins were prepared (mAbs 2F6, 2G8, 6E2, and 7E3). MAb 7E3, which had the highest binding affinity for the Cap protein, was chosen for further investigation. A novel B cell epitope DLDGAW was identified using mAb 7E3. An epitope-blocking (EB) enzyme-linked immunosorbent assay (ELISA) was successfully developed using horseradish-peroxidase-labeled mAb 7E3 to detect PCV3 antibodies in porcine sera. Moreover, the EB-ELISA showed no specific reaction with other porcine disease sera, and the cut-off value was defined as 35%. Compared with the commercial ELISA, the percentage agreement was 95.59%. Overall, we have developed a novel EB-ELISA method that accurately and conveniently detects PCV3 in serum, making it a valuable tool for the clinical detection of PCV3 infection.
猪圆环病毒3型（PCV3）在全球猪群中呈地方流行性，可引起繁殖障碍、皮炎和肾病综合征以及多器官炎症。目前，疾病监测中对快速准确的诊断方法的需求日益增长。在本研究中，制备了四种针对PCV3衣壳蛋白的单克隆抗体（mAb）（mAb 2F6、2G8、6E2和7E3）。选择对Cap蛋白具有最高结合亲和力的mAb 7E3进行进一步研究。使用mAb 7E3鉴定了一个新的B细胞表位DLDGAW。使用辣根过氧化物酶标记的mAb 7E3成功开发了一种表位阻断（EB）酶联免疫吸附测定（ELISA），用于检测猪血清中的PCV3抗体。此外，EB-ELISA与其他猪病血清无特异性反应，临界值定义为35%。与商业ELISA相比，一致性百分比为95.59%。总体而言，我们开发了一种新型EB-ELISA方法，可准确便捷地检测血清中的PCV3，使其成为临床检测PCV3感染的有价值工具。
