Mycobacterium avium ssp. paratuberculosis (MAP) is the cause of chronic gastroenteritis in cattle called the Johne's disease (JD). The disease causes significant economic losses in cattle production. MAP is also supposed to be involved in the Crohn's disease and inflammatory bowel disease (IBD) in people. The detection of the cattle infection based on investigations of milk samples and evaluation of the capacity of the methods used to detect the disease was the objective of the present study. Following methods were applied for milk samples testing: detection of MAP in bacterial culture, detection of the specific IS-900 fragment of MAP in the genetic material isolated directly and detection of MAP antibodies. The results obtained were compared with the "golden standard" results, i.e. the isolation of MAP from the faeces. PQStat-the program for diagnostic reliability estimation, was used for evaluation of the sensitivity, specificity and predictive value. The method based on detection of the specific IS-900 fragment of MAP in the genetic material isolated directly from milk samples was found to possess the highest sensitivity. Detection of anti-MAP antibodies on the other hand showed the lowest sensitivity. The method of detecting anti-MAP antibodies in milk was the most specific while detection of the IS-900 fragment in the genetic material was the least specific method. These results obtained may serve as a guide to choose the most appropriate method for diagnosis of MAP infections by milk sample testing.