Detection of lymphocystis disease virus infection to flounder gill cells in vitro by monoclonal antibodies

摘要

Flounder gill (FG) cells were used to isolate lymphocystis disease virus (LCDV) and two monoclonal antibodies (Mabs) (1A8 and 3G3) against LCDV were used to trace LCDV infection to FG cells. FG monolayer cells was inoculated with LCDV supernatant, obtained from lymphocystis cells of diseased flounder, Paralichthys olivaceus . LCDV infection was detected with Mabs employing immunocytochemical assay (ICA) and indirect immunofluorescence assay test (IIFAT) technique. Detected by IIFAT, they were specific for LCDV. The results of experimental infection illustrated that FG cells was sensitive to LCDV, and showed virus- infection positive detected by ICA. Cytopathic effect (CPE) occurred 1-2 days post inoculation (PI) , and half tissue culture infection dosage (TCID_(50)) of virus supernatant was 2~(2.57) per 40 μl. Tracing by IIFAT showed that LCDV positive signal first appeared at the cell membrane immediately PI , and then in cytoplasm at 24h PI , it reached the strongest positive at 48-72 h PI , and began to decrease at 96h PI.