In this study, twenty five bulk tank milk samples each of 50ml from Bulk tank milk of 25 holstein-friesian dairy herds representing 7 Egyptian Governorates, were collected at the period from January to July 2013. Each raw bulk tank milk sample was subjected to ELISA and real time PCR, while detection of viable Mycobacterium avium subsp. paratuberculosis (MAP) was done by culture of milk samples on Herrold's egg yolk media "HEYM". The results indicated that real time PCR detected 17out of 25 samples (68 %) followed by ELISA 10 samples out of 25 (40%), more over the viable MAP was isolated from 15 out of 25 samples (60%), the autors indicated the high percentage of MAP isolation may be to milk contamination with infected faeces and shedded microorganisms directly in milk. The authors concluded that, Since the milk is considered an essential nutrient for childrens and adults, it should be free of any hazard pathogens could be transmitted to it by direct or indirect way, so it is very important in milk value chain to test the raw bulk milk provided to plant milk processing by efficient and fast test as real time PCR to exclude any possible threat for the consumer by using the efficient pasteurization for that pathogen