Jul 12, 2022
Japanese encephalitis virus isolation
- Wenjing Liu1,
- Shihong Fu2
- 1Department of Basic Medicine, School of Medicine, Qingdao University, Qingdao, People’s Republic of China;
- 2Department of Arbovirus, NHC Key Laboratory of Biosafety, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, People’s Republic of China
Protocol Citation: Wenjing Liu, Shihong Fu 2022. Japanese encephalitis virus isolation. protocols.io https://dx.doi.org/10.17504/protocols.io.bdaui2ew
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it’s working
Created: March 06, 2020
Last Modified: July 12, 2022
Protocol Integer ID: 33844
Abstract
The mosquito grinding supernatant and CSF samples were inoculated into Vero cells to finish virus isolation.
Materials
The mosquito grinding supernatant ,CSF samples, Vero cell.
Before start
Mosquito grinding supernatant was sterilized using a filter with a pore size of 0.22 μm.
virus isolation
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 .