Cloning and characterization of two Onchocerca volvulus repeated DNA sequences

Two repeated sequences, plasmids pOV8 and pOV26, were cloned and characterized from the filarial parasite Onchocerca volvulus. Both clones can be used to distinguish O. volvulus DNA from other Onchocerca species or other nematodes by restriction fragment length polymorphisms, but neither clone can differentiate between DNA from savanna (Mali) or forest (Ivory Coast) O. volvulus isolates. DNA from one O. volvulus infective larva can be detected by both clones in dot blot hybridization assays. Neither clone cross hybridizes with DNA from host or vector species (human or simuliid, respectively). pOV26 is a member of an interspersed repeated DNA family composed of at least 100 members, and is only observed in the genus Onchocerca. Repeated DNA clone pOV8 cross reacts with DNA from other parasitic filarial nematodes, and is also present in at least 100 copies per O. volvulus genome. pOV26 is a potential tool in the diagnosis of human onchocerciasis, since it is specific for the genus Onchocerca. In the future, we plan to look for regions of these repeated sequences which may serve as a basis for the development of probes to discriminate among Onchocerca species and strains using a simple dot hybridization test.