Sun Shaohua, Zhang Kaili, Zhang Jiajia, Zhang Pingping, He Ping, Deng Dafu, Jiang Sen, Zheng Wanglong, Chen Nanhua, Bai Jianfa, Zhu Jianzhong
College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China.
Joint International Research Laboratory of Agriculture and Agri-Product Safety, Yangzhou University, Yangzhou 225009, China.
Int J Mol Sci. 2025 Mar 14;26(6):2619. doi: 10.3390/ijms26062619.
Porcine reproductive and respiratory syndrome (PRRS) is a pig respiratory disease threating the global swine industry. To combat PRRS, it is necessary of the effective diagnostic detection of antibody, including developing a neutralizing antibody against porcine reproductive and respiratory syndrome virus (PRRSV), especially the currently prevalent NADC30-like PRRSV in China. In this study, we prepared three monoclonal antibodies (mAbs) against NADC30-like PRRSV glycoprotein 5 (GP5) protein, and identified two corresponding precise epitopes (WR and QWGRP). In the neutralization test, QWGRP recognizing GP5 mAbs (11E6 and 12D1) exhibited obvious neutralizing activity, whereas the WR recognizing mAb (3A8) showed low neutralizing activity. Based on the two antigenic peptides, a peptide-based Enzyme-Linked Immunosorbent Assay (ELISA) was developed to detect antibodies against PRRSV, presenting high specificity, sensitivity, and repeatability. The concordance rate of the peptide-based ELISA and commercial IDEXX PRRSV X3 Ab ELISA in detection of 81 clinical samples was 82.7%. In conclusion, the GP5 peptide-based ELISA can be used for the detection of neutralizing antibodies against NADC30-like PRRSV, providing a rapid and reliable method for monitoring PRRSV infection.
猪繁殖与呼吸综合征（PRRS）是一种威胁全球养猪业的猪呼吸道疾病。为对抗PRRS，有效诊断检测抗体很有必要，包括研发针对猪繁殖与呼吸综合征病毒（PRRSV）的中和抗体，尤其是中国目前流行的NADC30样PRRSV。在本研究中，我们制备了三种针对NADC30样PRRSV糖蛋白5（GP5）蛋白的单克隆抗体（mAb），并鉴定出两个相应的精确表位（WR和QWGRP）。在中和试验中，识别GP5的QWGRP单克隆抗体（11E6和12D1）表现出明显的中和活性，而识别WR的单克隆抗体（3A8）表现出低中和活性。基于这两种抗原肽，开发了一种基于肽的酶联免疫吸附测定（ELISA）来检测抗PRRSV抗体，具有高特异性、敏感性和重复性。基于肽的ELISA与商业IDEXX PRRSV X3 Ab ELISA在检测81份临床样本中的符合率为82.7%。总之，基于GP5肽的ELISA可用于检测针对NADC30样PRRSV的中和抗体，为监测PRRSV感染提供了一种快速可靠的方法。
