Thirty HIV-1-positive samples from Bolivia were genetically characterized on the basis of HMA and DNA sequencing, revealing the presence of B and F subtypes, in accordance with the molecular epidemiology pattern already described for other South American countries such as Brazil and Argentina. The interpatient divergence of subtype B Bolivian specimens was on average 14.2% (4.3-19.8%) at the nucleotide level, whereas the two unlinked subtype F samples (BO23 and BO29) were only 8.2% divergent, suggesting a more recent introduction of this subtype in the country. In our study group, which represents 13% of the HIV/AIDS cases already described in Bolivia as of May 1996, the transmission occurred more frequently through heterosexual exposures (46.7%), followed by homosexual (23.3%), bisexual (10%), intravenous drug use (3.3%), and vertical (3.3%); in one case the potential exposure category could not be defined (3.3%). No association could be established between exposure categories, gender, or clinical classification and subtype distribution in the Bolivian HIV/AIDS patients.