Evaluation of the critical parameters of a sensitive ELISA test using purified infectious bovine rhinotracheitis virus antigens

David C. Bolton, Hsien Jue Chu, Alex Ardans, Barry Kelly, Chung Zee Yuan Chung Zee

Research output: Contribution to journalArticle

28 Scopus citations

Abstract

An enzyme-linked immunosorbent assay (ELISA) for the survey or titration of bovine sera for the presence of IgG antibodies against infectious bovine rhinotracheitis (IBR) virus was developed. The optimal conditions of serum dilution, antigen concentration, conjugate dilution, substrate concentrations, and reaction time were established using the signal/ noise (S/N) ratio as the determining criterion. Equilibrium density gradient purified IBR virus was used as antigen at an optimal concentration of 0.60 μg/cuvette. The use of purified antigen allowed the testing of sera at a 1 : 10 dilution without nonspecific reaction. The conditions of conjugate dilution, substrate concentration and reaction time were shown to have significant effects on the ELISA test. Results from 35 sera showed this optimized ELISA procedure to be as much as 1000-fold more sensitive than the serum neutralization plaque reduction assay. Numerous sera showing no neutralizing titer to IBR virus were found to be positive when examined by this ELISA method.

Original languageEnglish (US)
Pages (from-to)265-279
Number of pages15
JournalVeterinary Microbiology
Volume6
Issue number4
DOIs
StatePublished - Jan 1 1981

ASJC Scopus subject areas

  • Microbiology
  • veterinary(all)

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