We have investigated the cause of dual serological reactivity to human immunodeficiency virus (HIV) types 1 and 2, a common occurrence in West Africa. Serum specimens from 111 individuals from Côte d'Ivoire classified by commercial western blot as HIV-1 (n = 15), HIV-2 (32), and dually reactive (64) were further tested by more specific serological tests (a synthetic peptide enzyme immunoassay [Pepti-LAV 1/2] and western blots prepared from antigen in which oligomeric forms of the transmembrane protein were disrupted by trichloroacetic acid [WB-TCA]). Peripheral blood mononuclear cells were tested for HIV-1 and HIV-2 with the polymerase chain reaction (PCR) and virus culture. Of 104 samples that were concordant by both WB-TCA and Pepti-LAV, 82 (79%) were confirmed by PCR results. Virus culture was concordant with serology for specimens (35/38) in which any virus was detected. Our findings indicate that mixed HIV-1/HIV-2 infections are common in Côte d'Ivoire, and suggest that natural infection by one HIV type does not prevent heterologous infection.