Single-strand conformation polymorphism analysis for identification of four members of the Anopheles funestus (Diptera: Culicidae) group

Members of the Anopheles funestus Giles group are difficult to identify because of the morphological overlap that exists within the group. This inability to distinguish species, as well as the fact that the species vary in their behavior and biting preferences, complicate the successful planning and maintaining of malaria control programs. In this article we discuss the use of a single-strand conformation polymorphism (SSCP) assay to distinguish 4 members of the An. funestus group collected at 10 different localities in Africa. rDNA genes differ at numerous sites among closely related species. Using conserved primers, the D3 domain in the 28S gene was amplified electrophoresed on SSCP gels, and species-specific patterns were observed. Intraspecific variation was detected in An. funestus specimens from East and West Africa. Analyzing 108 An. funestus, 78 An. vaneedeni Gillies & Coetzee, 21 An. rivulorum Leeson, and 2 An. leesoni Evans, we concluded that SSCPs can be used successfully as a molecular tool for the identification of these species.