The aim of this study was the analysis of human immunodeficiency virus (HIV) diversity in Venezuela, and the characterization of variants other than subtype B. A total of 425 HIV isolates, collected between 2003 and 2008, were analyzed. The sequence of at least one genomic region (Pol, Env, Vif, or Nef ) was available for all of them and at least two genomic regions were analyzed in 46% of them. From the 425 HIV isolates analyzed, 421 (99.1%) were classified as HIV-1 subtype B. The four non-subtype B isolates correspond to one subtype C, one recombinant AG, and two HIV-2 isolates. This study shows that HIV-1 subtype B is still highly predominant in Venezuela. Whereas some sporadic cases of other HIV types can be found, they do not seem to have disseminated to the present.