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Schughart3 project protocol

Susceptibility to influenza A (H3N2) viral infection: body weight and survival at three doses in the 8 inbred founder strains of the Collaborative Cross   (2013)

Leist SR, Kollmus H, Schughart K




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

    1. H3N2 is produced in the allantoic cavity of 10-d-old embryonated hen eggs for 48h at 37°C.
    2. Mice are weighed to determine dosing.
    3. Mice are anesthetized by intra-peritoneal injection with a mixture of Ketamine/Xylazine in sterile natrium chloride solution.
    4. 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).
    5. 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.
    6. 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