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Clinical and Vaccine Immunology, March 2006, p. 395-402, Vol. 13, No. 3
1071-412X/06/$08.00+0     doi:10.1128/CVI.13.3.395-402.2006

Development and Application of Reference Antisera against 15 Hemagglutinin Subtypes of Influenza Virus by DNA Vaccination of Chickens

Chang-Won Lee,1 Dennis A. Senne,2 and David L. Suarez1*

Southeast Poultry Research Laboratory, USDA Agricultural Research Service, Athens, Georgia 30605,1 National Veterinary Services Laboratories, USDA Animal and Plant Health Inspection Service, Ames, Iowa 500102

Received 2 September 2005/ Returned for modification 5 December 2005/ Accepted 4 January 2006

Reference antisera were produced against 15 influenza hemagglutinin (HA) subtypes using DNA vaccination to produce a high-quality polyclonal serum to the HA protein without antibodies to other influenza viral proteins. The HA gene from each of 15 different HA subtypes of influenza virus was cloned into a eukaryotic expression vector and injected intramuscularly, together with a cationic lipid, into 3- to 4-week-old specific-pathogen-free chickens. Birds were boostered twice at 4-week intervals after the initial injection, and in general, antibody titers increased after each boost. The antisera were successfully applied in the hemagglutination inhibition test, which is the standard method for the classification of the HA subtypes of influenza virus. We also demonstrated the HA specificity of the antisera by Western blot and immunodot blot analysis. DNA vaccination also provides a safer alternative for the production of HA-specific antibodies, since it is produced without the use of live virus.

* Corresponding author. Mailing address: Southeast Poultry Research Laboratory, USDA-ARS, 934 College Station Road, Athens, GA 30605. Phone: (706) 546-3479. Fax: (706) 546-3161. E-mail: dsuarez{at}seprl.usda.gov.

Clinical and Vaccine Immunology, March 2006, p. 395-402, Vol. 13, No. 3
1071-412X/06/$08.00+0     doi:10.1128/CVI.13.3.395-402.2006




This article has been cited by other articles:

  • Chen, M.-W., Cheng, T.-J. R., Huang, Y., Jan, J.-T., Ma, S.-H., Yu, A. L., Wong, C.-H., Ho, D. D. (2008). A consensus-hemagglutinin-based DNA vaccine that protects mice against divergent H5N1 influenza viruses. Proc. Natl. Acad. Sci. USA 105: 13538-13543 [Abstract] [Full Text]  


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