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Purpose: To evaluate the safety and efficacy of highly active antiretroviral therapy (HAART) in human immunodeficiency virus (HIV) infected pregnant women to prevent mother-to-child transmission of HIV. Design of the study is observational retrospective evaluation of a prospective cohort. Setting of the study is King Chulalongkorn Memorial Hospital. Participants are HIV-infected Thai women and their offspring. HIV-positive pregnant women who received antenatal care were given antepartum and intrapartum zidovudine (ZDV) plus single dose nevirapine (NVP) or HAART [ZDV + lamivudine (3TC) + NVP] to prevent HIV transmission to the neonate. All newborns received single dose NVP plus 6-weeks of ZDV.HAART was given to their parents if indicated. We followed the infants from February 2003 to June 2006 to determine HIV-status. Calculated HIV maternal-child transmission rate is the main outcome. Among 157 mother-infant pairs, 44 mothers received ZDV plus single dose NVP and 113 mothers received HAART. There was one HIV-infected child in the first group and 3 in the second group, resulting in 2.3% (95% confidence interval, −2.1 to 6.7) transmission rate in the first group versus 2.7% (95% confidence interval, −0.3 to 5.6) in the second group (P = 1.000). All non-infected infants had a normal physical examination at 18 months. Initiation of HAART in HIV-infected pregnant women had comparable efficacy with ZDV plus single dose NVP to prevent mother-to-child transmission of HIV. In addition, the program may have strengthened the role of the family unit and stimulated public awareness reproductive health planning can help in prevention of HIV/AIDS.