Sun Shaohua, Zhang Kaili, Zhang Jiajia, He Ping, Zhang Pingping, Deng Dafu, Chi Chenglin, Jiang Sen, Zheng Wanglong, Chen Nanhua, 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; Comparative Medicine Research Institute, Yangzhou University, Yangzhou 225009, China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou 225009, China.
Vet Microbiol. 2025 Mar;302:110399. doi: 10.1016/j.vetmic.2025.110399. Epub 2025 Jan 20.
Porcine reproductive and respiratory syndrome virus (PRRSV) causes reproductive failure in sows and respiratory disease in growing pigs, leading to significant economic losses worldwide. Due to the constant mutation and recombination, PRRSV exhibits significant genetic diversity, the general detection of all PRRSV-2 and PRRSV-1 strains is thus needed. In our study, four monoclonal antibodies (mAbs) against PRRSV nucleocapsid (N) protein were generated and the precise and novel B cell epitopes (KPHF and HHTVR) were identified. The epitope KPHF is highly conserved across all strains of PRRSV-2 lineages and PRRSV-1 subtypes, and the corresponding two mAbs (6D7, 4D12) were selected to develop a sandwich ELISA that was able to detect all tested PRRSV-2 and PRRSV-1 strains. The developed sandwich ELISA demonstrated high specificity, sensitivity and repeatability. In detection of 133 clinical samples, the sandwich ELISA achieved 84.21 % coincidence with the real-time RT-PCR. In conclusion, the mAb based sandwich ELISA can be suitable for detection of potential all PRRSV-2 lineages and PRRSV-1 subtypes, providing a simple, quick and high content method for diagnosis of PRRS.
猪繁殖与呼吸综合征病毒（PRRSV）可导致母猪繁殖失败以及生长猪的呼吸道疾病，在全球范围内造成重大经济损失。由于不断发生突变和重组，PRRSV呈现出显著的遗传多样性，因此需要对所有PRRSV - 2和PRRSV - 1毒株进行常规检测。在我们的研究中，制备了四种针对PRRSV核衣壳（N）蛋白的单克隆抗体（mAb），并鉴定出了精确且新颖的B细胞表位（KPHF和HHTVR）。表位KPHF在所有PRRSV - 2谱系和PRRSV - 1亚型毒株中高度保守，选择相应的两种单克隆抗体（6D7、4D12）开发了一种夹心ELISA，该方法能够检测所有测试的PRRSV - 2和PRRSV - 1毒株。所开发的夹心ELISA表现出高特异性、敏感性和重复性。在检测133份临床样本时，夹心ELISA与实时RT - PCR的符合率达到84.21%。总之，基于单克隆抗体的夹心ELISA适用于检测所有潜在的PRRSV - 2谱系和PRRSV - 1亚型，为PRRS的诊断提供了一种简单、快速且含量高的方法。
