On-site differential diagnostic detection of HP-PRRSV and C-PRRSV using EuNPs-mAb fluorescent probe-based immunoassay
ANALYTICAL AND BIOANALYTICAL CHEMISTRY(2021)
Abstract
Porcine reproductive and respiratory syndrome virus (PRRSV) has caused worldwide economic losses in the swine industry. Pigs infected with highly pathogenic (HP)-PRRSV display more severe symptoms than those infected with classical (C)-PRRSV. A rapid, sensitive, and reliable detection method to distinguish between HP-PRRSV and C-PRRSV is needed. In this study, we prepared a monoclonal antibody from a hybridoma that can distinguish HP-PRRSV(including TP, QJ, LQ, JN-HS, and TY strain) from C-PRRSV (CH-1A strain) using cell surface-fluorescence immunosorbent assays (CSFIA). Based on this monoclonal antibody (4D5), we developed a europium microsphere-based lateral flow immunochromatographic strip (EuNPs-LFICS) for the differential diagnostic detection of HP-PRRSV and C-PRRSV. Under optimized conditions, the method was rapid (15 min), sensitive (LOD: 2.57 ng mL −1 , 606 TCID50/0.1 mL), selective for HP-PRRSV detection, and quantitative (DLR: 3.56–228 ng mL −1 ). In clinical samples, the EuNPs-LFICS assay was largely consistent with PCR results, indicating its practical clinical application. Graphical abstract
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Key words
Porcine reproductive and respiratory syndrome virus, Monoclonal antibodies, Molecular modeling, Europium microspheres, Differential diagnosis
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