Affiliation:
1. Southeast Poultry Research Laboratory, Agricultural Research Service, U.S. Department of Agriculture
2. Department of Infectious Diseases, College of Veterinary Medicine, University of Georgia, Athens, Georgia
Abstract
ABSTRACT
Vaccination programs for the control of avian influenza (AI) in poultry have limitations due to the problem of differentiating between vaccinated and virus-infected birds. We have used NS1, the conserved nonstructural protein of influenza A virus, as a differential diagnostic marker for influenza virus infection. Experimentally infected poultry were evaluated for the ability to induce antibodies reactive to NS1 recombinant protein produced in
Escherichia coli
or to chemically synthesized NS1 peptides. Immune sera were obtained from chickens and turkeys inoculated with live AI virus, inactivated purified vaccines, or inactivated commercial vaccines. Seroconversion to positivity for antibodies to the NS1 protein was achieved in birds experimentally infected with multiple subtypes of influenza A virus, as determined by enzyme-linked immunosorbent assay (ELISA) and Western blot analysis. In contrast, animals inoculated with inactivated gradient-purified vaccines had no seroconversion to positivity for antibodies to the NS1 protein, and animals vaccinated with commercial vaccines had low, but detectable, levels of NS1 antibodies. The use of a second ELISA with diluted sera identified a diagnostic test that results in seropositivity for antibodies to the NS1 protein only in infected birds. For the field application phase of this study, serum samples were collected from vaccinated and infected poultry, diluted, and screened for anti-NS1 antibodies. Field sera from poultry that received commercial AI vaccines were found to possess antibodies against AI virus, as measured by the standard agar gel precipitin (AGP) test, but they were negative by the NS1 ELISA. Conversely, diluted field sera from AI-infected poultry were positive for both AGP and NS1 antibodies. These results demonstrate the potential benefit of a simple, specific ELISA for anti-NS1 antibodies that may have diagnostic value for the poultry industries.
Publisher
American Society for Microbiology
Reference44 articles.
1. Alexander, D. J. 2000. A review of avian influenza in different bird species. Vet. Microbiol.74:3-13.
2. Beard, C. W. 1970. Avian influenza antibody detection by immunodiffusion. Avian Dis.14:337-341.
3. Beard C. W. 1989. Serological procedures p. 192-200. In H. Friedman (ed.) A laboratory manual for the isolation and identification of avian pathotypes. Kendall-Hunt Publishing Dubuque Iowa.
4. Birch-Machin, I., A. Rowan, J. Pick, J. Mumford, and M. Binns. 1997. Expression of the nonstructural protein NS1 of equine influenza A virus: detection of anti-NS1 antibody in post infection equine sera. J. Virol. Methods65:255-263.
5. Buonagurio, D. A., S. Nakada, J. D. Parvin, M. Krystal, P. Palese, and W. M. Fitch. 1986. Evolution of human influenza A viruses over 50 years: rapid, uniform rate of change in NS gene. Science232:980-982.
Cited by
88 articles.
订阅此论文施引文献
订阅此论文施引文献,注册后可以免费订阅5篇论文的施引文献,订阅后可以查看论文全部施引文献