Identification of Anopheles parensis (Diptera: Culicidae) using ribosomal DNA internal transcribed spacer (ITS2) sequence variation

Anopheles funestus Giles (Diptera: Culicidae) is arguably the most efficient vector of malaria in Africa. It belongs to a group of nine morphologically similar species, most of which play no role in malaria transmission. Studies on the An. funestus group are hampered by the difficulties in identifying members of the group. A polymerase chain reaction single-strand conformation polymorphism (PCR-SSCP) assay can accurately distinguish three species within the group, namely An. funestus Giles, An. rivulorum Leeson and An. leesoni Evans. However, the SSCP gel profiles of two additional species, An. parensis Gillies and An. vaneedeni Gillies & Coetzee, although different from the above three species, overlap in their profiles, leading to misidentification. We report on the development of an additional PCR assay able to discriminate between An. vaneedeni and M. parensis on the basis of fragment size variation after amplification of the ITS2 region of rDNA. Combining these two assays it is possible to identify five of the most common species of the An. funestus group found in southern Africa.