Highly sensitive antigen detection procedures for the diagnosis of infectious bovine rhinotracheitis: amplified ELISA and reverse passive haemagglutination.

The sensitivity of an enzyme-linked immunosorbent assay (ELISA) for the detection of bovid herpesvirus 1 antigen was increased by up to 50-fold using the biotin-avidin interaction to amplify the reaction, when compared with a simple sandwich ELISA. An alternative immunoassay, reverse passive haemagglutination (RPHA), had a similar sensitivity to the amplified ELISA, and was technically simpler to perform. Both the amplified ELISA and the RPHA could detect viral antigen in the nasal secretions of calves undergoing experimental primary infection with the virus from Day 3 to Day 7 after inoculation. Neither assay was as sensitive as virus isolation in cell culture and they failed to detect antigen in virus-positive samples from the calves from 8 days after inoculation, and from vaccinated calves undergoing challenge infection.