+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

Observations on the preparation and stability of infectious bronchitis virus hemagglutination antigen from virus propagated in chicken embryos and chicken kidney cell cultures



Observations on the preparation and stability of infectious bronchitis virus hemagglutination antigen from virus propagated in chicken embryos and chicken kidney cell cultures



Avian Diseases 28(2): 504-513



A total of 166 infectious bronchitis virus (IBV) hemagglutination (HA) antigen preparations were made during a 30-month period from allanto-amnionic fluid (AAF) from chicken embryos inoculated with 10 different IBV strains (Mass 41, Conn 46, H52, Florida 18288, Ark 99, JMK, T, Holte, EF, SE17). Antigens were prepared by inoculating 9- or 10-day-old embryos with 10(5.0) to 10(6.5) EID50 IBV, harvesting AAF after a 30-hour-postinoculation incubation, and phospholipase C (PLC) treatment of virus concentrated by pelleting from the AAF. Longer (48 hr) incubation times were tried, but production of H52 HA antigen was successful only from AAF harvested after 30 hours of incubation. AAF from JMK-infected embryos had lower infectivity titers and frequently yielded lower HA antigen titers than the other strains. The treatment of AAF with fluorocarbon did not enhance or diminish HA activity but did yield clearer antigens by removing extraneous material. Polyethylene glycol precipitation of virus was an acceptable alternative to pelleting virus at 39,000 X g. Inactivation of IBV with 0.1% betapropiolactone did not affect HA activity, whereas inactivation with 0.1% formalin caused a marked reduction in HA titer. Different buffer formulations including phosphate, tris, or HEPES were tried to optimize the conditions for PLC treatment of virus concentrate, but there were no apparent differences in the antigens prepared in the different buffers. The HA antigen preparations were stored and were stable at 4 C. Antigen titers of greater than or equal to 64 after storage for 20 months or longer were not uncommon. Addition of merthiolate as a preservative had no deleterious effect on HA activity. Antigen stability appeared to be enhanced by incorporating EDTA in buffer for virus pellet recovery and during enzyme treatment. Attempts to produce HA antigens from cell-culture-adapted virus propagated in chicken kidney cells were less satisfactory. An acceptable HA antigen was prepared from only two (Mass 41, SE17) of the seven strains that were tried. Virus propagation in chicken embryos is the better method of the two for IBV HA antigen production. Aside from the need to concentrate virus and treat the concentrate with PLC, there appeared to be considerable latitude in the procedures that can be used to make acceptable IBV HA antigens.

Please choose payment method:






(PDF emailed within 0-6 h: $19.90)

Accession: 043784340

Download citation: RISBibTeXText

PMID: 6331366

DOI: 10.2307/1590358


Related references

Comparative sensitivities of embryonating chicken's eggs and primary chicken embryo kidney and liver cell cultures to infectious bronchitis virus. Avian Dis 9(2): 308-316, 1965

Plaque formation by infectious bronchitis virus in chicken embryo kidney cell cultures. Avian Diseases 17(2): 369-378, 1973

Comparative sensitivities of oviduct and tracheal organ cultures and chicken embryo kidney cell cultures to infectious bronchitis virus. Avian Diseases 27(3): 594-601, 1983

Immune response in chickens in infectious bronchitis virus, strain 33. I. Response to beta-propiolactone-inactivated virus. II. Response to virus propagated in chicken embryo kidney cells. Avian Doseases 15(4): 688-695; 969-703, 1971

Immune response in chickens to infectious bronchitis virus, strain 33. II. Response to virus propagated in chicken embryo kidney cells. Avian Diseases 15(4): 696-703, 1971

Stabilizing effect of magnesium sulfate on avian infectious bronchitis virus propagated in chicken embryo kidney cells. Applied Microbiology 23(2): 281-284, 1972

Preparation and protective efficacy of a chicken embryo kidney cell-attenuation GI-19/QX-like avian infectious bronchitis virus vaccine. Vaccine 36(28): 4087-4094, 2018

Immunofluorescence of avian infectious bronchitis virus in primary chicken embryo kidney, liver, lung, and fibroblast cell cultures. Archiv für die Gesamte Virusforschung 19(3): 265-272, 1966

Concentration of infectious bronchitis virus antigen in chicken embryos for agar gel diffusion technique. Indian Journal of Animal Health 13(1): 71-72, 1974

Infectious bronchitis in the chicken. I. Virus cultivation in cultures of chick embryo kidney cells. Zentralblatt für Veterinarmedizin. Reihe B. Journal of Veterinary Medicine. Series B 13(4): 345-363, 1966

Research Note: Avian Encephalitis Virus as a Chicken Kidney Cell-Culture Contaminant and Its Effect on the Growth of Infectious Bronchitis Virus. Avian Diseases 17(3): 666-670, 1973

Research note--avian encephalitis virus as a chicken kidney cell-culture contaminant and its effect on the growth of infectious bronchitis virus. Avian Diseases 17(3): 666-670, 1973

The susceptibility of chicken kidney and oviduct organ cultures to a vaccine strain of avian infectious bronchitis virus. Research in Veterinary Science 26(1): 38-40, 1979

Coronavirus infectious bronchitis virus virus retaining spike glycopolypeptide s 2 but not s 1 is unable to induce virus neutralizing or hemagglutination inhibiting antibody or induce chicken tracheal protection. Journal of General Virology 67(7): 1435-1442, 1986

Purification of infectious bronchitis virus propagated in embryonated chicken eggs and its confirmation by RT-PCR. Indian Journal of Comparative Microbiology, Immunology and Infectious Diseases 24(2): 143-147, 2003