Thirty-two sheep with clinical paratuberculosis and 43 normal, healthy control animals were tested for serum antibodies to Mycobacterium avium subspecies paratuberculosis with the absorbed enzyme-linked immunosorbent assay (ELISA) and agar gel immunodiffusion (AGID) test. All the sheep were necropsied and the diseased cases were categorised as having either multibacillary (‘lepromatous’) or paucibacillary (‘tuberculoid’) intestinal lesions. The ELISA and AGID test were highly sensitive when testing the multibacillary group (86.4 per cent and 100 per cent, respectively) but the sensitivity of the tests in the paucibacillary group was significantly lower (10 to 50 per cent and 30 per cent, respectively). These findings were related to the ELISA optical density readings, with the multibacillary samples having values significantly greater than those of the paucibacillary and control groups, and the optical density values appearing to correlate with the numbers of mycobacteria present in the intestinal lesions. These results indicate both the usefulness of serological testing in the diagnosis of the multibacillary form of paratuberculosis and the difficulty in identifying animals with the paucibacillary form of the disease.