Japanese encephalitis virus isolation
Wenjing Liu, Shihong Fu
Abstract
The mosquito grinding supernatant and CSF samples were inoculated into Vero cells to finish virus isolation.
Before start
Mosquito grinding supernatant was sterilized using a filter with a pore size of 0.22 μm.
Steps
virus isolation
The filtered sterile mosquito grinding solution was inoculated into 24-well plates grown to 80% monolayer Vero cells, 70 μl of each well was added for virus isolation, and a 3-well blank control was set for each plate.(for CSF samples 100μl of each well was added for virus isolation).
Incubate at 37℃ for 1 h.
1 ml of MEM medium containing 2% FBS was added to each well for maintenance of growth. Cultivate at 37℃ 5% CO2 and observe under microscope every day whether cytopathic effect (CPE) occurs.
Specimens with no CPE were used for blind transmission for three generations .
