Boon1 project protocol

Susceptibility to influenza A (H5N1) viral infection in females of 23 strains of mice   (2011)

Boon AC, Webby RJ
With: Finkelstein D, Zheng M, Liao G, Allard J, Klumpp K, Webster R, Peltz G




Boon1_Protocol

Project protocol - Contents

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

Reagents and solutions

  • phosphate-buffered saline (PBS)

Procedure: H5N1 preparation and inoculation

    1. 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.
    2. Mice are inoculated intranasally in a volume of 30 µL PBS; EID50 six doses ranging serially from 10 to 1,000,000.
    3. 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

Data collected by investigator

  • body weight, pre-infection
  • *body weight, post-infection, days 4, 7, 10
  • percent changes in body weight, post infection
  • survival
  • *mortality

* Available in Supplementary data file only Boon1_suppdata.xlxs