Sequences encoding the envelope glycoprotein of human immunodeficiency virus type 1 (HIV-1) were amplified by PCR from plasma and peripheral blood mononuclear cells obtained at four time points from an acute seroconvertor. Genetic analyses, including nucleotide sequencing and heteroduplex mobility studies, showed that the patient harbored three distinct populations of HIV-1 clade B envelope sequences, with nucleotide distances ranging from 9.2 to 17.2%. One population of sequences was clearly distinguishable from the others on the basis of phylogenetic analysis. In addition, sequences suggesting recombination between two of the three distinct viral populations were also found. This case of acute seroconversion provides clear and conclusive evidence that coinfection by multiple HIV-1 strains can indeed occur in vivo.