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This study aims to evaluate the epidemiological characteristics of mother-to-child transmission (MTCT) of HIV and identify the possible factors leading to infant HIV infection using a retrospective cohort study of early infant diagnosis (EID). Information on a total of 3,145 exposed infant-mother pairs was collected from the EID platform from July 2014 to December 2019. The MTCT rate was 2.1%. Spatial-temporal maps showed that rates varied by year and by region, with four districts (Baise, Guigang, Guilin, and Hechi) maintaining rates of2.0% in 2019. The rate of antiretroviral therapy (ART) use was 94.4%, with a gradual increase in prescriptions of highly active ART (HAART) from 83.0% in 2014 to 92.4% in 2019. A majority of 99.5% of infants were receiving artificial feeding. Factors associated with MTCT were ART use (odds ratio [OR] = 0.065, confidence interval [95% CI] = 0.035-0.121) and artificial feeding (OR = 0.091, 95% CI = 0.018-0.452). HAART was more helpful in decreasing the risk of MTCT compared with monotherapy (OR = 0.115, 95% CI = 0.014-0.933). ART during the postpartum period correlated with an increased risk (OR = 11.579, 95% CI = 1.402-95.960) compared with use of ART during pregnancy. This study indicates that MTCT rate of HIV is decreasing meaningfully in Guangxi. Some areas still face challenges in elimination of MTCT and need further resources and interventions. Future program planning should take into consideration the fact that ART use-in particular the use of HAART or ART during pregnancy-and replacement feeding may contribute to the prevention of MTCT.