In this study 96 randomly selected fecal samples of two-year old or older cattle from Trakya district of Turkey were investigated for the presence of Mycobacterium avium subsp. Paratuberculosis by PCR. A pair of genus specific primers, derived from the IS 900 gene of the agent was used as a target sequence for the PCR. The specific DNA was not detected in any of the 96 fecal samples and all of them were considered to be PCR negative. In conclusion, although the absence of specific DNA in the samples of cattle raised in the Trakya districts is an optimistic sign, further studies on John's disease are needed.