Evaluation of a New Enzyme-Linked Immunosorbent Assay (ELISA) for Johne's Disease in Goat Serum

Abstract

Control of Johne’s Disease (JD) is a high priority, it causes significant economic losses in the livestock industry. An efficacious vaccine for JD is not currently available and no satisfactory treatment exists, so control programs depend on diagnostic tests based on detection of the immune response generated after infection with Mycobacterium avium subsp. paratuberculosis (MAP)-the causative agent of JD-to identify and remove infected animals and management practices to interrupt transmission. Objectives: The objective of this study was to evaluate a new, commercially-available, Enzyme-Linked, Immunosorbent Assay (ELISA) that was developed for cattle, for use with goat serum. For comparison, a United States Department of Agriculture (USDA)-approved ELISA kit for goats, an Agar Gel Immunodiffusion (AGID) test and a Polymerase Chain Reaction (PCR) test to detect MAP in fecal samples were also evaluated. Methods: Infection status at necropsy was taken as the reference standard, determined by gross, histological and microbiological examination of tissue specimens. The serum and fecal samples were taken from goat kids that had participated in a previous vaccine efficacy study. Eighty goat kids were vaccinated either once subcutaneously at eight weeks with a commercial vaccine, or orally at eight and 10 weeks with one of five experimental oral vaccines or a sham-control oral vaccine. Kids were challenged orally with a bovine isolate of MAP, three weeks after the last vaccination. Results: None of the vaccines prevented infection. Samples taken from 10 to 13 months post-challenge were used to compare the assays. The specificity of all tests was 100%. At 13 months, the new ELISA, approved ELISA, AGID test and fecal PCR test had sensitivities of 55.7%, 52.9%, 24.3% and 87.1% respectively. At 11 months the sensitivities were 48.6%, 38.6%, 10.0% and 78.6% respectively and the sensitivity of the new ELISA was significantly better than the approved ELISA (p = 0.0196). Conclusion: The new ELISA effectively detected circulating MAP antibody in the serum of infected goats and the preliminary results in this study indicate the kit has potential for use in goat JD control programs. The results for the new ELISA in goat serum are comparable with those reported when the ELISA test is used for cattle serum.