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Development of 316v antibody enzyme-linked immunosorbent assay for detection of paratuberculosis in sheep

Author(s) : R.B. Gurung, D.J. Begg, A.C. Purdie, G.J. Eamens & R.J. Whittington

Summary :

An enzyme-linked immunosorbent assay (ELISA) was developed and optimised using a Mycobacterium avium subspecies paratuberculosis (MAP) antigen prepared from a C strain (316v) passed through a French press. The optimised assay was evaluated with a panel of sera from MAP infected (n = 66) and uninfected (n = 1,092) sheep. Animals in the MAP infected category were positive on either tissue culture or histopathology but were of unknown serum antibody status. The diagnostic performance and cost of the assay were compared with those of a commercial ELISA (IDEXX). At 99.8% diagnostic specificity the assay showed a diagnostic sensitivity of 23% (95% CI: 15.1–35.8) compared with 36.4% (95% CI: 25.8–48.4) for the commercial ELISA (McNemar’s test: chi-square 5.82, p < 0.05). The sensitivities were 5.9% (95% CI: 1–26.9), 27.9% (95% CI: 14.7–45.7) and 35% (95% CI: 18.1–56.7), for low grade, paucibacillary and multibacillary lesion grades, respectively.


Antibody – Diagnosis – Paratuberculosis – Sensitivity – Serum – Sheep – Specificity.

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