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Sendai Virus Propagation Using Chicken Eggs

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Sendai Virus Propagation Using Chicken Eggs

Narihito Tatsumoto et al. Bio Protoc.

Abstract

Sendai virus is a member of the family Paramyxoviridae, and an enveloped virus with a negative-stranded RNA genome. Sendai virus is not pathogenic to humans, but for mice and can cause pneumonia in mice. Easy and efficient techniques for propagating Sendai virus are required for studying virus replication, virus-induced innate- and adaptive-immunity, Sendai-virus-based virotherapy and IgA nephropathy. Here, we describe a protocol for Sendai virus propagation using chicken eggs. This traditional protocol enables us to generate a large amount of virus enough for animal experiments as well as cell culture experiments in a relatively inexpensive way.

Keywords: Allantoic fluid; Chicken eggs; Harvest; Mouse parainfluenza virus; Propagation; Sendai virus.

Conflict of interest statement

Competing interests Authors declare no conflicts of interest or competing interests.

Figures

Figure 1.
Figure 1.. Chicken egg with air cell.
Air cell is visualized by candling (left). Draw a line along the edge of air cell (arrows in left panel) and mark X (right) in the position of the embryo with a pencil under a strong light in dark room. Note: This is an example of regular/unembryonated egg. You will see an embryo as a slightly dark spot and red vessels (right) by candling in the embryonated egg on Day 10.
Figure 2.
Figure 2.. Chicken egg with a puncture.
Make a hole with a push pin at the location 10 mm below the air cell edge and opposite to embryo.
Figure 3.
Figure 3.. How to open up an egg.
A. Making a hole on the shell over air cell. B. Cutting shell by scissors along but slightly inside of the line of air cell. C. Exposed air cell and intact shell membrane. D. Peeling and removal of shell membrane with forceps completely; E. Collecting allantoic fluid between shell membrane and an embryo (yolk in this picture) with an 18 G needle and 50 ml syringe. Note: This is an example of unembryonated egg. You will see an embryo instead of yellow yolk in the actual viral propagation.
Figure 4.
Figure 4.. Collected allantoic fluid.
Allantoic fluid is transparent or slightly cloudy with yellowish tinge.

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