关键词: 仙台病毒抗原, 酶联免疫吸附试验, 免疫印迹试验, 间接免疫荧光试验

Abstract: Abstract: Objective To prepare and purify the murine Sendai virus ( SeV) antigen so as to establish the indirect ELISA and Western blot test for detecting the antibody against SeV． Methods SeV was inoculate into the allantoic cavity of SPFchick embryo，and then the allantoic fluid was added to the BHK-21 cell culture to proliferate SeV． Sendai virus antigen was prepared and purified by the differential centrifugation and ultrasonication，and its purity was identified by SDS-PAGE． The best coating antigen concentration for the indirect SeV-ELISA was selected and confirmed． SeV antibody in murine serum was screened by the indirect immunofluorescence assay ( IFA) ，and the SeV-ELISA specificity was verified by Western-blot test． Results The HA titer of Sendai Virus from BHK-21 cell culture was 1∶ 128，and SeV antigen electrophoretic purity reached to 80 percent． While the best coating antigen concentration for the indirect SeV-ELISA was assigned to 2. 5 μg /mL，the detection rate of positive Sev antibody in murine serum by the SeV-ELISA was absolutely compatible with that by IFA． Conclusions The SeV antigen prepared by differential centrifugation and sonication broken method can be used for detection of Sendai Virus antibody in mice，and the antigen-specific and activity of Sendai Virus have been verified by the Western-blot test．

Key words: Sendai Virus antigen, ELISA, Western blot, IFA