Identifying the socioeconomic and structural issues that act as enablers and/or barriers to HIV testing services is critical in combatting HIV/AIDS amongst mothers and children in Africa. In this study, we used a weighted sample of 46,645 women aged 15-49 who gave birth in the two years preceding the survey from the recent DHS dataset of ten East African countries. Multivariable logistic regression was used to investigate the factors associated with prenatal HIV test uptake in East Africa. The overall prenatal HIV test uptake for the prevention of mother-to-child transmission (PMTCT) of HIV was 80.8% (95% CI: 74.5-78.9%) in East Africa, with highest in Rwanda (97.9%, 95% CI: 97.2-98.3%) and lowest in Comoros (17.0%, 95% CI: 13.9-20.7%). Common factors associated with prenatal HIV test service uptake were higher maternal education level (AOR = 1.29; 95% CI: 1.10-1.50 for primary education and AOR = 1.96; 95% CI: 1.53-2.51 for secondary or higher education), higher partner education level (AOR = 1.24; 95% CI: 1.06-1.45 for primary education and AOR = 1.56; 95% CI: 1.26-1.94 for secondary or higher school), women from higher household wealth index (AOR = 1.29; 95% CI: 1.11-1.50 for middle wealth index; AOR = 1.57; 95% CL: 1.17-2.11 for rich wealth index), improved maternal exposure to the media, and increased awareness about MTCT of HIV. However, residents living in rural communities (AOR = 0.66; 95% CI: 0.51-0.85) and travelling long distances to the health facility (AOR = 0.8; 95% CI: 0.69-0.91) were associated with non-use of prenatal HIV test service in East African countries. In each East African country, factors associated with prenatal HIV test uptake for PMTCT varied. In conclusion, the pooled prenatal HIV test uptake for PMTCT of HIV was low in East Africa compared to the global target. Scaling up interventions to improve enablers whilst addressing barriers to the use of prenatal HIV test services are essential to end the HIV/AIDS epidemic in East African countries.