Project
protocol
-
Contents
Workflow
and
sampling
Step |
Procedure |
Data
collected |
1 |
Mouse-adapted H3N2 produced and prepared for intra-nasal delivery |
- |
| 2 |
Mice weighed to determine weight changes |
Body weight |
| 3 |
Mice anesthetized |
- |
| 4 |
Mice infected with H3N2 intra-nasally |
- |
| 5 |
Body weight determined daily post-infection |
Body weight |
| 6 |
Mice monitored daily and survival recorded |
Survival, susceptibility score |
Equipment
and
supplies
- Incubator
- Balance
- Syringes and needles
Reagents
and
solutions
- Ketamine/xylazine (100 mg/mL and 20 mg/mL, respectively)
- H3N2: mouse-adapted virus strain influenza A/HK/01/68 obtained from Otto Haller, University of Freiburg GERMANY
- Natrium chloride solution
- Phosphate-buffered saline
Procedure: H3N2 preparation/delivery and daily monitoring of infected mice
- H3N2 is produced in the allantoic cavity of 10-d-old embryonated hen eggs for 48h at 37°C.
- Mice are weighed to determine dosing.
- Mice are anesthetized by intra-peritoneal injection with a mixture of Ketamine/Xylazine in sterile natrium chloride solution.
- Mice are intra-nasally infected with 20 µL virus solution of sterile phospate-buffered saline using a concentration of 10 focus-forming units (FFU), 2000 FFU, or 200,000 FFU (3 cohorts, each cohort getting a different dose).
- Body weight is measured daily and determined as percentage of initial weight at day 0 (pre-infection); mice that lose more than 30% of their body weight over the course of the experiment are euthanized and scored as dead.
- Survival is recorded.
Data
collected
by
investigator
- Body weight pre-infection
- Body weight post-infection (expressed as percentage of pre-infection weight)
- Survival
- Susceptibility score
|