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Evaluation of the abilities of three diagnostic tests based on the polymerase chain reaction to detect Mycobacterium paratuberculosis in cattle: application in a control program.

Department of Bacteriology, School of Veterinary Medicine, University of Utrecht, The Netherlands.

ABSTRACT

Three assays for the specific detection of Mycobacterium paratuberculosis by dot spot hybridization of polymerase chain reaction products were applied to fecal samples of dairy cattle. The first two tests used polymerase chain reaction primers and a DNA probe derived from M. paratuberculosis-specific sequences of the 16S rRNA gene and insertion element IS900, respectively. These two tests were carried out on spiked fecal samples to determine the detection limits. The 16S rRNA test was able to detect 10(7) bacteria per g of feces, and the IS900 test detected 10(4) to 10(5) per g of feces. Next, we studied the usefulness of these tests in a control program for paratuberculosis. Therefore, the tests and a third, commercially available, test (IDEXX Corp.) were used twice with an interval of 3 months on fecal samples of 87 cows from two dairy herds with a history of Johne's disease. We compared the results of these tests with those of culturing. This showed that the tests are specific but that the sensitivity ranged from 3 to 23%. Further improvement of the sensitivity is needed before the tests can be used in a control program to eradicate Johne's disease.

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