A Novel Peptide-Based Enzyme-Linked Immunosorbent Assay (ELISA) for Detection of Neutralizing Antibodies Against NADC30-like PRRSV GP5 Protein

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 (¹⁵⁵WR¹⁵⁶ and ¹⁹⁶QWGRP²⁰⁰). In the neutralization test, ¹⁹⁶QWGRP²⁰⁰ recognizing GP5 mAbs (11E6 and 12D1) exhibited obvious neutralizing activity, whereas the ¹⁵⁵WR¹⁵⁶ 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.