动物造模 z-bio 2022-11-21 350

　　Objective: To establish the infection model of EV71 on primary renal cells of tree shrews.

　　Methods: The primary renal cells of tree shrews were obtained by trypsin digestion. The renal cells of tree shrews were infected with EV71. The viral titers of the supernatants were measured at 1,2,4,6 and 8 days after culture. The expression of EV71 virus VP1 protein in the cells was detected by Western blot and indirect immunofluorescence, respectively, to determine the infectivity of EV71 virus to the primary renal cells of tree shrews.

　　Results: The isolated primary kidney cells of tree shrew were subcultured and purified, and their morphology was identified. The primary kidney cells of tree shrews were infected with EV71 virus, and the virus titer reached 1.3 48~96 h after infection × 106 TCID50/mL, indicating that EV71 virus can effectively infect primary kidney cells of tree shrew and effectively proliferate. Western blot showed that EV71 virus VP1 protein could be effectively detected in the primary kidney cells of tree shrews 2~8 days after infection, and the distribution of virus VP1 protein was detected in the cytoplasm of cells 2~6 days after infection by indirect immunofluorescence method.

　　Conclusion: Based on the successful establishment of primary renal cell culture of tree shrew, the infectivity and viral proliferation characteristics of EV71 virus on primary renal cells of tree shrew were determined, and the primary renal cell infection model of EV71 tree shrew was preliminarily established.