HIV/AIDS is currently a major public health problem in Ethiopia and mother to child transmission (MTCT]. The transmission of HIV from infected mothers to babies could occur during antenatal period, as well as during delivery and breastfeeding (postnatal period). Since breastfeeding is essential for child survival, it is also necessary to assess mothers’ knowledge attitude and practice towards HIV transmission and its prevention during breastfeeding. Objective of the study is to assess knowledge attitudes and practice of pregnant women attending Durame general hospital regarding PMTCT of HIV infection. A cross sectional, descriptive study was conducted to assess knowledge attitude and practice towards PMTCT among ANC clients in Durame general hospital from November 28, 2018 to June 20, 2018. A systematic random sampling technique was used to select 200 ANC attendants. Data was analyzed using tally sheet, and finally presented with table and graphs. 220 pregnant women were participated in the study which accounts (100%). 215 of the respondents reported that they have heard about HIV/AIDS. 165(75%) know that MTCT of HIV is preventable. 210(95.5%) of them were willing to have HIV test and the reasons mentioned were to protect their partner and child. Finally, it is concluded that most of the ANC clients knew HIV transmissions from infected mothers to their child during pregnancy, delivery and breast feeding and it is preventable.