Background: Prevention of mother to child transmission of Human Immunodeficiency Virus (PMTCT) programme requires great male participation to attain Ethiopia’s new plan to elimination of mother to child transmission of Human Immunodeficiency Virus by 2015. In this regard, antenatal care is one of the areas where male couple is expected to participate through voluntary counselling and testing. Currently, studies showed that male involvement and support in the program is critical to improving women’s uptake of the services. Objectives: To assesses male involvement in prevention of mother to child transmission of HIV in the context to partner testing and associated factors at antenatal care in Goba town, Bale zone, and Oromia region. Methods: A facility-based cross-sectional study was conducted. A total of 422 male partners, whose spouses were attending antenatal care at Goba hospital were enrolled through systematic sampling method. The study was also supplemented by two focus group discussions. The data were cleaned and coded then analyzed using SPSS for windows version 16. Odds ratio were computed to assess the presence and degree of association between dependent and independent variables. Logistic regression analysis was carried out to see independent effect of each variable on the outcome. Qualitative data was also analyzed by thematic content. Result: Only 22.7% of male partners accompanied their spouses for HIV testing and counselling at antenatal clinic. Men having no formal education was 90% less likely to be tested as compared to those who had completed the higher education [AOR=0.1, 95%CI: 0.023-0.050, P=0.004]. Identified barriers to male involvement in the PMTCT programme were individual factors such as being busy, proxy testing, fear of being tested and neglecting importance, inadequate knowledge about the program, health system factors and absence of legally enforced law that makes male to participate. Conclusion and Recommendation: Majority of male partners had not participated in prevention of mother to child transmission (PMTCT) program at antenatal clinic. To promote uptake of the service by male partner, there is a need to raise awareness of the program among community through behavioral change and communication, develop a new strategy for promoting male participation in the program and couple HIV testing laws should be revised to get HIV free generation.