The Study Of Virion Attachment Proteins (VAP) Of WSSV By Monoclonal Antibodies

White spot syndrome virus (WSSV) emerged in 1993 and caused severe damage to shrimp culture industry. Therefore, WSSV has been extensively studied, resulting in some exciting research progress. It is believed that the binding of virion attachment protein (VAP) on the envelope with receptors plays important roles in virus infection, so it is important to study the VAP and receptors. We primarily cultured haemocytes of Fenneropenaeus chinensis, infected with WSSV, and detected the WSSV and its propagation in cultured cells. Enzyme linked immunosorbent assay (ELISA), DOT-BLOT, primary cultured haemocytes and crayfish (Cambarus proclarkii) were used to detect neutralizing effect of monoclonal antibodies (Mabs) against WSSV. We also purified envelope protein of WSSV, produced Mab againt VP19, and detect the neutralization of the Mab.Hemocytes were withdrawn from heart of F. chinensis, cultured with seven kinds of culture medium, the hemocytes remained viable for about 20 days using leibovitz's 2×L-15 medium consisting of 20% FBS, 20% GIT, and 2% glucose, adjusting osmolarity with sodium chloride, pH 7.2-7.4, and incubating at a temperature of 25℃.Different concentrations of virus inoculums inoculated the primary cultured hemocytes, the infected cells were observed with an inverted phase-contrast microscope, and found the cytopathic effects of infected cells were diverse. 10^-1-10^-6 concentrations of WSSV inoculums inoculated primary cultured hemocytes of F. chinensis, cell death was detected by phase-contrast microscope for 5d. Hemocytes cultures were infected with 10^-3 concentration of virus inoculums for different times and with 10^-1-10^-8 concentrations of virus inoculums for 10d, respectively, virus in hemocytes were detected by ELISA with mixture of monoclonal antibodies against WSSV and HRP linked antibodies. The result showed that hemocytes which were infected with 10^-1-10^-2 concentrations of virus inoculums shrinked, cell death and cell debris were found, as time going on, the number of hemocytes declined, the cell debris piled; Hemocytes that were infected with 10^-3-10^-4 concentrations of virus inoculums...