DAO 143:195-203 (2021)  -  DOI: https://doi.org/10.3354/dao03561

ABSTRACT: Cyprinid herpesvirus 2 (CyHV-2) has become a serious threat to the gibel carp Carassius auratus gibelio industry and has led to enormous losses worldwide. We have therefore developed an immunochromatographic strip (ICS) to enable rapid on-site detection of CyHV-2 by aquaculture facility staff. The ICS employs 2 monoclonal antibodies (MAbs 2C3-1E6 and 3H2-1G5) against the ORF25 protein, a CyHV-2 membrane protein, as the capture and detection antibodies, respectively. Indirect immunofluorescence assay (IIFA) and Western blotting of CyHV-2-infected fathead minnow cells indicated that the 2 MAbs could specifically bind CyHV-2 by recognizing ORF25 antigen. Sandwich ELISA showed that the detection limit of ORF25 protein halved when MAb 2C3-1E6 served as the capture antibody compared to MAb 3H2-1G5. The test for detecting purified CyHV-2 using the ICS could be completed in 10 min and the sensitivity was 1 µg ml^-1. Sensitivity of the ICS remained stable following storage at 4, 25 and 37°C for 6 mo. Tissue homogenate from gibel carp with and without obvious gill hemorrhages was subjected to CyHV-2 detection using the ICS: the results were in good accordance with conventional PCR. Our ICS does not require highly trained technicians or specialized equipment, making it suitable for rapid diagnosis of CyHV-2 infection both in the laboratory and in the field.

KEY WORDS: Immunochromatographic strip · Cyprinid herpesvirus 2 · ORF25 · Monoclonal antibodies · Rapid diagnosis · Gibel carp · Carassius auratus gibelio