The aim of this study was to investigate the implementation of a prevention of mother-to-child transmission (PMTCT) program and to evaluate the uptake and adherence to single-dose nevirapine in a cohort sample that had undergone PMTCT in five public clinics in a resource-poor setting, Quakeni Local Service Area, O.R. Tambo District in the Eastern Cape, South Africa. Results indicated that 116 women (15.3% of the sample) were infected with HIV, 642 (84.7%) were uninfected, and 552 (42.1%) had an unknown HIV status. Almost all of the women had received information about HIV and HIV testing prenatally, but 552 (42%) had not been tested for HIV, and their HIV status was unknown. Only 66 (57%) of the HIV-infected pregnant women had been provided with nevirapine. It is recommended that the quality of HIV counseling be improved and the program of maternal self-medication with nevirapine tablets at onset of labor and maternal provision of nevirapine syrup to newborns be encouraged.