Abstract:Selecting small molecular ligands or monoclonal antibodies to cellular receptors as inhibitors is an effective approach to inhibit virus infection. In this research, haemocyte plasma membranes (HPM) of Chinese shrimp (Fenneropenaeus chinensis) were isolated employing differential centrifugation. Utilizing the techniques of Dot-Blot and ELISA, HPMs immobilized in microtiter plats or nitrocellulose (NC) membranes were incubated with Digoxingenin labeled white spot syndrome virus (WSSV-DIG) at 4℃ for 4 h. Then alkaline phosphatase labeled antibodies against DIG was used as a sensitive probe to detect the in vitro combination of WSSV-DIG to HPM. Both ELISA and Dot-Blot combination showed positive reactions. In blocking experiment, polyclonal antibody against haemocytes of Chinese shrimp was allowed to bind to the HPM at 37℃ for 1 h before the incubation with WSSV-DIG. In Dot-Blot assay, the blocked group showed a significantly lighter color compared with the unblocked group. In ELISA assay, OD value of the blocked group was 69% lower than the unblocked group. It can be deduced that some receptor antibodies, which were specifically against the WSSV receptors, existed in the polyclonal antibody. During incubation, these antibodies adhered to the receptors on the HPM and significantly blocked the combination of WSSV-DIG. The in vitro combining technology between WSSV and HPM developed in this research provide a strategy to characterization of WSSV receptors and selection inhibitor to WSSV. The blocking experiments carried out with polyclonal antibody have proved the feasibility.