Triatoma dimidiata Latreille is a major vector of Chagas disease with an extensive geographic distribution from Central Mexico, through Central America, to northern South America. As a result of its variability in phenetic and genetic characters, disagreement concerning its taxonomic status has been raised. In this study, the cuticular hydrocarbon pattern of T. dimidiata populations from Mexico, Belize, Guatemala, Honduras, Costa Rica, and Colombia was analyzed by capillary gas chromatography coupled to mass spectrometry; linear discriminant analysis was used to help elucidate population structure. Vector populations segregated into five distinct groups; specimens from Yucatan Peninsula, together with those from Central Mexico, Central America, and Colombia corresponded to different T. dimidiata subspecies, a putative different species comprising insects from Belize, together with an isolated population collected at bat caves in Guatemala. The analysis revalidates the earlier division of T dimidiata into three subspecies, T. d. maculipennis, T. d. dimidiata, and T. d. capitata; and an additional subspecies and a distinct species are proposed.