Preparing Hemagglutinating Antigen from Isolates of Infectious Bronchitis Virus

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Abstract
The hemagglutinating (HA) activity of 14 strains of infectious bronchitis virus (IBV) was investigated. The optimal conditions for IBV antigen preparation include inoculation of 10 or 11 day old specific-pathogen-free embryonated eggs and incubation for 30 h at 37.degree. C. Embryos were inoculated via the allantoic cavity with 0.1 ml of a low embryonic passage of the virus (107-108 median egg infectious doses/ml). Allantoic fluid was harvested and pooled and a 100-fold concentration of virus particles was achieved by centrifugation for 3 h at 30,000 .times. g. Virus pellets were resuspended in Tris-hydrochloride buffer containing 3 U of phospholipase C (type 1)/ml and incubated for 2 h at 37.degree. C. All IBV strains tested demonstrated positive HA activity with chicken red blood cells. The antigen was stored in liquid state or lyophilized at 4.degree. C. [This HA antigen may be suitable as a diagnostic reagent for use in monitoring chickens for IBV by the hemagglutination-inhibition test.].

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