BACKGROUND: The scourge of Human Immunodeficiency Virus/Acquired Immune Deficiency Syndrome (HIV/AIDS) worldwide is unprecedented with a disproportionately higher prevalence in sub-Saharan Africa. HIV in pregnancy is of particular importance given the risk of mother-to-child transmission. It is even more important giving the fact that a previously tested HIV negative pregnant woman can seroconvert or acquire new infection in the course of pregnancy thus increasing the burden of this disease in our region. An assessment of the prevalence of HIV seropositivity in labour and corresponding infant HIV infection will allow appropriate intervention to be carried out and more children will be saved from being infected with HIV, otherwise eradication of Paediatric HIV will remain a mirage. AIM AND OBJECTIVES: This study determined the prevalence of HIV seropositivity in labour among all booked pregnant women who earlier tested negative in antenatal clinic of Imo State University Teaching Hospital Orlu and the corresponding rate of mother-to-child transmission. METHODOLOGY: This was a prospective observational study of all booked pregnant women in labour who earlier tested negative in the antenatal HIV routine testing and counseling (RTC) at booking conducted over 4 months. The patients found to be non-reactive at booking had repeat HIV screening done in labour with serial rapid test algorithm. For any seropositive patient during the repeat testing, the baby HIV testing by DNA polymerase chain reaction was done at six week postpartum. All the subjects also filled questionnaires. Data analysis was done using SPSS version 20.The confidence interval was 95%.