Contagious agalactiae (C.A.) is one of the most common disease affecting small ruminants which is caused by Mycoplasma agalactiae. This disease is particularly widespread around the world and Iran is one of the countries that C.A. is present. The aim of this study was isolation and identification of M. agalactiae (MG) with culture and PCR technique in milk samples in Kordestan province, Iran. A total of 367 milk samples were collected from sheep and goat. Specific published primers amplify a 375 bp gene of MG were used for PCR. Twenty (5.5%) out of 367 were positive in PPLO agar and 5 (25%) out of these isolates were positive with Mycoplasma agalactiae primers. Four (75%) out of 5 isolates was from sheep and 1(25%) from goat. Result of PCR with 367 milk samples showed that 11(3%) of them were positive with these primers. The isolation of M. agalactiae showed that C.A is present in Kordestan province and our results suggested that PCR method because of reduces the time consuming could be an alternative method beside culture.