Project protocol - Contents
- Workflow and sampling
- Reagents and solutions
- Procedure: H5N1 preparation and inoculation
- Definitions and calculations
Workflow and sampling
Step Procedure performed *Data collected 1 Prepare viral inoculum - 2 Weigh mice (pre-infection) - 3 Inoculate mice 4 Monitor mice 30 days for morbidity and mortality; weigh mice periodically body weight, survival
* Additional data available in Boon1_suppdata.xlxs
Procedure: H5N1 preparation and inoculation
- A reverse genetics variant of A/HongKong/213/03, a highly pathogenic H5N1 influenza A virus, is propatated in 10-day-old embryonated chicken eggs as described in Boon et al., 2011.
- Mice are inoculated intranasally in a volume of 30 µL PBS; EID50 six doses ranging serially from 10 to 1,000,000.
- Mice are monitored for 30 days for signs of morbidity and mortality; body weights and survival are recorded.
Definitions and calculations
- EID: egg infective dose
- EID50 lo: 10, 100, 1000 (for grouping data)
- EID50 med: 10,000 (for grouping data)
- EID50 hi: 100,000 to 1,000,000 (for grouping data)
- Resistance/susceptibility: investigators set cut off at 104 EID50
- body weight, pre-infection
- *body weight, post-infection, days 4, 7, 10
- percent changes in body weight, post infection
* Available in Supplementary data file only Boon1_suppdata.xlxs