Propagation and Harvest of Influenza Virus from a Fertilized Chicken Egg

Abstract

Source: Brauer, R. et al., Influenza Virus Propagation in Embryonated Chicken Eggs. J. Vis. Exp. (2015)

This video demonstrates the synthesis and harvest of the influenza virus in fertilized chicken eggs. The virus is inoculated into the allantoic cavity, where it replicates in epithelial cells and buds into the allantoic fluid, which is then collected and stored for downstream applications.

Protocol

1. Influenza Virus Inoculation via the Allantoic Route
   1. On the day of virus inoculation (day 10 or 11), candled the eggs again and eliminated any dead embryos. Distinguish live embryos by looking for movement inside the egg.
   2. Remove three eggs from the incubator, and arrange the eggs in a separate holder with the air sac up.
   3. Mark the air space of the eggs with a permanent marker.
   4. Wash the eggshell above the air space with 70% ethanol.
   5. Put the eggs with the air space up in an egg carton and into a biosafety hood (biosafety level 2 or BSL-2).
   6. Use a sterile 18-G needle to punch a small hole in th....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Serum pathogen-free (SPF) fertilized chicken eggs	VALO BioMedia
Humidified egg incubator	FARM iNNOVATORS	Model 2100
Automatic egg turner	FARM iNNOVATORS	Model 3200
Egg candler	FARM iNNOVATORS	Model 3300
1 ml Syringe	BD Bioscience	309659
18G Needle	BD Bioscience	305196
Ethanol>99.5%	Sigma-Aldrich	459844	Diluted to 70% using water
Glue gun "Ad tech Hi Temp Project Pro"	Ad tech, Adhesive Technologies, Inc.	Model 1105
Glue "Ad tech MINI SIZE, Multi Temp"	Ad tech, Adhesive Technologies, Inc.	220-34ZIP30