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Diseases of Aquatic Organisms

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

Development and evaluation of a convenient immunochromatographic strip test for rapid detection of cyprinid herpesvirus 2 (CyHV-2)

Ronghua Wu1,2,3,#, Yang Xue4,#, Jianan Huang2, Eda Ozdemir3, Yun Li1, Shihua Ding1,*

1College of Fisheries, Southwest University, Chongqing 400715, PR China
2Key Laboratory of Freshwater Fish Reproduction and Development, Minister of Education, Southwest University, Chongqing 400715, PR China
3Department of Basic Sciences, College of Veterinary Medicine, Mississippi State University, Mississippi State, MS 39762, USA
4Chongqing Fisheries Technical Extension Center, Chongqing 400020, PR China
*Corresponding author:
#These authors contributed equally to this work

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


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Cite this article as: Wu R, Xue Y, Huang J, Ozdemir E, Li Y, Ding S (2021) Development and evaluation of a convenient immunochromatographic strip test for rapid detection of cyprinid herpesvirus 2 (CyHV-2). Dis Aquat Org 143:195-203. https://doi.org/10.3354/dao03561

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