动物造模,EV71病毒 z-bio 2022-01-14 456

　　OBJECTIVE: To establish an EV71 virus infection model of tree shrew primary renal epithelial cells.

　　Methods: The primary kidney cells of tree sh were obtained by trypsin digestion, and the kidney cells of tree sh were infected with EV71. Western blot and indirect immunofluorescence were used to determine 1, 2, 4, and 6 respectively, and the culture was measured at 8°C. Virus titer of the supernatant. In each cell, the expression of EV71 virus VP1 protein was detected, and the infectivity of EV71 virus to shrimp primary kidney cells was determined.

　　Result: The isolated primary kidney cells of the tree sh were passaged, purified, and morphologically identified, and a cell culture of the primary kidney cells based on the tree sh was established. After the EV71 virus was infected in the primary kidney cells infected by the tree, the virus titer reached 1.3×106 TCID50/mL 48-96 hours after infection, and the EV71 virus was infected in the primary kidney cells infected by the tree, indicating that it can Was effectively infected. And spread it effectively. Westernblot detection can effectively detect EV71 virus VP1 protein in tree primary kidney cells 2-8 days after infection, and indirectly detect the distribution of virus VP1 protein in the cytoplasm 2-6 days after infection. Immunofluorescence.

　　Conclusion: Based on the successful establishment of Tree Schrew primary kidney cell culture, the infectivity and viral growth characteristics of EV71 virus to Tree Schrew primary kidney cells were determined, and the EV71 Tree Schrew primary kidney cell infection model was determined.