Mycobacterium avium subspecies paratuberculosis causes Johne\\\'s disease which is widely distributed and is responsible for considerable economic losses throughout the world. There are a variety of diagnostic methods for molecular detection of the disease with PCR as one of the fastest, easiest, most sensitive and most specific tests. However, the efficiency of the conventional PCR has been hampered by the presence of small amounts of bacterium and considerable amounts of PCR-inhibitory factors in feces. To overcome these problems and in order to increase the limit of detection of the causing bacteria in feces, we developed a nested-PCR based on the conserved regions of IS900 genomic element. This sensitive test is able to accurately identify Mycobacterium avium subspecies Paratuberculosis. However to be able to validate this test and use it in the national prevention and control programs, its results need to be rigorously compared with the results of the standard methods.

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