Development and Evaluation of an ELISA for the Detection of Antibody to Infectious Laryngotracheitis Virus in Chickens

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Abstract
An enzyme-linked immunosorbent assay (ELISA) for the detection of antibody to infectious laryngotrachetis (ILT) virus in chickens was developed and compared with the serum-neutralization assay. The ELISA routinely yielded 16- to 32-fold higher titers than the serum-neutralization test. To overcome the requirement for large amounts of purified viral antigen, the microtiter trays were initially coated with an antibody prepared against purified ILT virus. A relatively crude viral preparation could then be used to coat the trays. Sera from specific-pathogen-free chickens < 12 wk of age did not show nonspecific binding, although 2.7% of all sera from chickens 13-64 wk of age had nonspecific activity. The majority of nonspecific reactors came from 1 highly inbred flock of specific-pathogen-free chickens. A number of modifications of ELISA procedures reported to reduce the nonspecific binding of chicken sera was investigated. Treatment of the serum or the plate and changes in the composition of the diluent did not increase the relative sensitivity of the anti-ILT assay.