动物造模,-EV71病毒,原代肾上皮细胞感染模型 z-bio 2021-04-26 255

　　Objective: To establish a tree shrew primary kidney cell EV71 infection model.

　　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 Treeshrew primary kidney cells were passaged, purified, and morphologically identified, and a cell culture based on Treeshrew primary kidney cells was established. When the EV71 virus is infected with primary kidney cells of the infected tree, the virus titer reaches 1.3×106 TCID50/mL 48-96 hours after infection. The EV71 virus infects the primary kidney cells infected by the tree, indicating that it can be effectively infected . 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.

　　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.