African reed frogs are widespread and highly polymorphic for colour pattern. The Hyperolius viridiflavus complex includes 121 described taxa, including subspecies, which are presently synonymised as 16 species. However, the taxonomy is confused as colour patterns have been used to identify species, despite an inadequate understanding of pattern variation and overlap. This study examined 267 sequences of the 16S rRNA fragment from individuals across the range of the group, from Senegal to Ethiopia, and to the southern tip of Africa. Maximum likelihood analysis showed that there were 14 species that only partially agreed with existing taxonomy. This study recognised the following species: Hyperolius viridiflavus, H. marmoratus, H. marginatus, H. parallelus, H. nitidulus, H. glandicolor, H. tuberculatus, H. mariae, H. noblei, H. goetzei, H. spatzi, H. bangwae, H. dintelmanni and an undescribed species from southern Cameroon. Hyperolius swynnertoni was found to be embedded within H. marmoratus, H. hutsebauti was embedded within H. parallelus, H. rhodesianus and H. pyrrhodictyon were embedded within H. marginatus and H. sheldricki was embedded within H. glandicolor. There was no sequence information to determine the status of Hyperolius nimbae. The ranges of the species are presented, although these are expected to change as more sequences become available. The use of DNA sequences to identify animals is recommended in this group, as the variation in advertisement calls and morphology is not well understood.