Hpall endonuclease distinguishes between two species in the Anopheles funestus group

The Anopheles funestus group consists of at least eight species that are currently identified mainly on morphological criteria. Until recently, only An. funestus s.s. was implicated in the transmission of malaria in Africa, but recent work in Tanzania has shown that An. rivulorum is also involved, albeit to a lesser degree than An. funestus. The constraints in the identification of the species and the need to clarify better their epidemiological role have led to the development of a PCR-RFLP method for the identification of two anthropophilic members of the group. Using PCR primers developed from the D3 region in the 28S ribosomal gene, amplified products were digested with the restriction endonuclease Hpall. This produced two distinct fragments on an agarose gel that could be used to separate An. funestus from An. vaneedeni. The technique needs to be tested on natural populations of these two species as well as on other members of the An. funestus group.