© The Author(s) 2016. With support from the U.S. President’s Emergency Plan for AIDS Relief and the global fund for HIV, tuberculosis, and malaria, Nigeria offers free services for prevention of mother-to-child transmission of HIV. However, uptake of these services is low, and pediatric transmission of HIV remains a significant public health challenge. Using the PEN-3 cultural model as the theoretical framework, we examined social, cultural, and contextual factors that influenced uptake of HIV counseling and testing among pregnant women and their male partners. This was a qualitative study of participants in the Healthy Beginning Initiative (HBI), a congregation-based program to prevent mother-to-child transmission of HIV in Enugu, southeast Nigeria. We conducted eight focus group discussion sessions with 83 pregnant women and their male partners. Participants’ perspectives on why they did or did not test for HIV were obtained. The most cited reasons for getting tested for HIV included the following: “the need to know one’s status”, “the role of prenatal testing” (positive perceptions); “the role of the church”, “personal rapport with healthcare worker” (positive enablers); and the “influence of marriage” (positive nurturer). The most cited reason for not testing were: “fear of HIV test”, “shame associated with HIV+ test results”, “conspiratorial beliefs about HIV testing” (negative perceptions); “lack of confidentiality with HIV testing”, (negative enabler); and “HIV-related stigma from family and community systems” (negative nurturer). Overall, numerous facilitators and barriers influence uptake of HIV testing in the study setting. Public health practitioners and policymakers need to consider how sociocultural and religious factors unique to specific local contexts may promote or hinder uptake of available HIV/AIDS prevention and care interventions.