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Use of faecal quantitative PCR testing to detect Johne's disease in sheep

S. Kawaji, D. J. Begg, K. M. Plain, R. J. Whittington

JOHNE'S disease, caused by Mycobacterium avium subspecies paratuberculosis (MAP), has significant economic impacts worldwide. This study evaluated the ability of a previously developed direct faecal real-time quantitative PCR assay to detect MAP infection in sheep.

Fifty-eight Merino sheep were included in the study. A total of 38 sheep were inoculated orally with MAP on three occasions when the animals were four months old. Twenty control sheep were kept separately from the inoculated animals throughout the study. All animals were weighed monthly; animals that had lost more than 10 per cent bodyweight over a one-month period were classified as clinically affected. Faecal and serum samples were collected before, and at four, eight and 13 months following inoculation. Postmortem examinations were also conducted at 13 months. Faecal samples were cultured and tested by PCR, serum samples were tested using a commercial ELISA, and intestinal tissue samples were cultured for the detection of MAP.

Ten of 38 inoculated animals developed clinical disease. At four months following inoculation, 18 of 38 sheep were detected by PCR to …

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