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Diseases of Aquatic Organisms

    DAO prepress abstract   -  DOI: https://doi.org/10.3354/dao03561

    Development and evaluation of a convenient immunochromatographic strip test for rapid detection of cyprinid herpesvirus 2

    Ronghua Wu, Yang Xue, Jianan Huang, Eda Ozdemir, Yun Li, Shihua Ding*

    *Corresponding author:

    ABSTRACT: The immunochromatographic strip (ICS) allows on-site rapid detection of cyprinid herpesvirus 2 (CyHV-2) without the requirement of specialized equipment and professional personnel, which is conducive to prevention and control of CyHV-2 infection. The ICS employed two monoclonal antibodies (MAbs) against ORF25 protein, a CyHV-2 membrane protein, as the capture and detection antibody, respectively. Ascites MAbs 2C3-1E6 and 3H2-1G5 were produced and purified, with anti-CyHV-2 titres of 1:2,048,000 and 1:1,024,000, respectively. Indirect immunofluorescence assay (IIFA) and Western blot of CyHV-2 infected FHM cells indicated that the two MAbs could specifically bind CyHV-2 by recognizing ORF25 antigen. Sandwich enzyme-linked immunosorbent assay (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. Sensitivity of the ICS remained stable following storage at 4°C, 25°C and 37°C for 6 months. Tissue homogenate from gibel carp (Carassius auratus gibelio) with and without obvious gill hemorrhages was subjected to CyHV-2 detection using the ICS: the results were in good accordance with conventional PCR. The developed ICS was highly specific and sensitive and could be completed within 10 min without the need for well-trained technicians and special equipment, making it suitable for rapid diagnosis of CyHV-2 infection both in the laboratory and field.