Samples from infants infected in-utero by the human immunodeficiency virus type 1 (HIV-1) subtypes A, C, D, and recombinants from Dar es Salaam, Tanzania, were examined for the presence of viral genetic quasispecies. HIV-1 envelope diversity was measured on peripheral blood mononuclear cells collected within the first 48 h of life from 53 infants. Phylogenetic analysis of C2-C5 envelope nucleotide sequences was used for HIV-1 subtype classification. Forty-two of 53 samples (79%) showed a heteroduplex mobility assay (HMA) suggestive of transmission of a single quasispecies, while 21% showed infection with multiple quasispecies. No differences among HIV-1 subtypes were found in the proportion of single to multiple quasispecies transmitted in-utero (Likelihood ratio test, P = 0.83), nor were differences found among single to multiple quasispecies transmitted and maternal viral load (Mann-Whitney test, P = 0.44). This suggests that differences in perinatal transmission between subtypes we previously observed in this cohort could not be associated with the likelihood for multiple independent infections during in-utero infections.