The polymerase chain reaction and cultural method was applied to detect Mycobacterium avium subspecies paratuberculosis (M. p.) in fecal samples of 165 suspected and of 35 diseased cattle, 18 intestinal tissues and 14 lymph nodes of diseased ruminants as well as organ material (n = 10) were also tested. An agreement (+/+, -/-) in the results between both methods was found in 89.7% of all cases examined. 12.7% of samples of suspected and 9.1% of diseased animals were only positive by PCR. In cases of intestinal lymph node and tissue as well as udder tissue of diseased cattle a total agreement between PCR and culture was observed. Lymph node of lung of one diseased cow was positive only in PCR.