A systematic review of genera and subgenera of Siphonaptera Rhopalopsyllinae comprising nine operating taxonomic units is presented based on phenetic and cladistic analysis. Twenty-seven characters of adults were analyzed and the character states were determined for each character by the out-group method. Phenogram and cladograms indicated two tribes, each with two subtribes. Systematic changes proposed as a result of this study include the raising of the subgeneric status of Gephyropsylla, Hechteilla, and Ayeshaepsylla to independent genera; the recognition of a new genus, Neotropsylla, to include Polygenis guimaraesi Linardi; the division of the genus Polygenis Jordan into two subgenera: Neopolygenis subgen. n., to include species of the atopus group, and Polygenis s. str., for remaining species.