Specific detection of M. avium subsp. paratuberculosis infections by EVELISA in calves experimentally inoculated with M. avium subsp. paratuberculosis (MAP), M. avium subsp. avium (MAA), and M. bovis (MB). Serum samples were obtained from calves intratonsillarly inoculated with M. avium subsp. paratuberculosis (n = 3; squares, calf 5903; circles, calf 5902; triangles, calf 5904), Mycobacterium avium subsp. avium (n = 3; squares, calf 193; circles, calf 6137; triangles, calf 2016), M. bovis (n = 3; squares, calf 202; circles, calf 2354; triangles, calf 203) and saline (control; n = 2; squares, calf 191; triangles, calf 192) at 1- or 2-week intervals for up to 321 days. A cutoff value of 0.24 (dotted line) was used to distinguish M. avium subsp. paratuberculosis-negative and -positive results. Similar results were obtained in two separate experiments.