Optimization of parameters for detecting antibodies against infectious bronchitis virus using an enzyme-linked immunosorbent assay: temporal response to vaccination and challenge with live virus.

James Case, Alex Ardans, D. C. Bolton, B. J. Reynolds

Research output: Contribution to journalArticle

21 Scopus citations

Abstract

Critical parameters affecting sensitivity and specificity of an enzyme-linked immunosorbent assay (ELISA) for avian infectious bronchitis virus (IBV) were evaluated and optimized. The use of purified IBV as antigen at 50 ng protein/well and high-ionic-strength serum dilution buffer has resulted in a test with minimal nonspecific binding of chicken immunoglobulins and very high sensitivity. Optimum conditions for serum dilution, conjugate dilution, and substrate incubation were determined for minimizing background and nonspecific reactions. The use of this test in a controlled challenge study with chickens vaccinated with live IBV demonstrated its effectiveness in monitoring circulating antibody levels to infectious bronchitis. The IBV ELISA, which is rapid, inexpensive, highly sensitive, and capable of handling very large numbers of samples, should provide the poultry industry with a reliable means for IBV flock monitoring.

Original languageEnglish (US)
Pages (from-to)196-210
Number of pages15
JournalAvian Diseases
Volume27
Issue number1
DOIs
StatePublished - Jan 1 1983
Externally publishedYes

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ASJC Scopus subject areas

  • Food Animals
  • Animal Science and Zoology
  • Immunology and Microbiology(all)

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