Preparation and Application of Monoclonal Antibodies against ORF72 of Cyprinid Herpesvirus 2
XU Chaonan1,2, SUN Yuyu2, WANG Jia1,2, GUO Baoqin1,2, WEI Chang2, LI Qiang2
1. Key Laboratory of Mariculture & Stock Enhancement in North China's Sea, Ministry of Agriculture and Rural Affairs, Dalian Ocean University, Dalian 116023, China; 2. Department of Ocean Technology, College of Marine and Biology Engineering, Yancheng Institute of Technology, Yancheng 224051, China
Abstract：In order to establish an immune detection method based on antigen levels, the nucleocapsid protein ORF72 of cyprinid herpesvirus Ⅱ (CyHV-2) was prepared by prokaryotic expression method. The purified ORF72 protein was immunized to BALB/c mice, and the anti-ORF72 monoclonal antibodies designated as 1B3, 4B1, 4C2, 5F4 and 5H9 were finally screened by hybridoma technology. Western blot analysis showed that the monoclonal antibodies specifically recognized recombinant protein ORF72 in vitro. The monoclonal antibody 4C2 detected the virus infection tissues of gibel crucian carp Carassius auratus gibelio by indirect immun of luorescence assay (IFA) only. IFA of the CyHV-2 replication on RyuF-2 detection with 4C2 showed that the positive signals were detected 36 hours after the virus infection, and the expression level of ORF72 protein was increased in the cell with time. Monoclonal antibody 4C2 contributes to research the replication mechanism of CyHV-2 in vitro and to establishment of the immune detection method for diseased fish.
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2024, Vol. 43 
