A study was conducted to determine the role of members of the Anopheles funestus group inmalaria transmission in the Mpumalanga Province, in the northeastern region of South Africa. Female anophelinemosquitoes were collected between January 1996 and November 1991 by means of human landing catches andtested for salivary gland, Plasmodium falciparum infections by means of the enzyme-linked immunosorbentassay (ELISA) method with PF2AIO antibodies. Infection rates from April and May 1997 collections were3.'737o and 19.4o/o, respectively. None of the nonimmune collectors became infected with malaria. The ELISApositivemosquitoes were tested with a polymerase chain reaction (PCR) malaria detection assay based onsequence variation present in the small subunit ribosomal RNA gene. Only l.O97o of EllSA-positive mosquitoeswere PCR-positive for malaria. Initially, all mosquitoes were assumed to belong to the An. .funestus group butsubsequent molecular taxonomy showed this assumption to be false. The use of a single-strand conformationpolymorphism (SSCP) assay revealed only 1 member of the An. funestus group, An. rivulorum. All other specimensproduced banding patterns not seen before. Those samples were identified morphologically as An. demeilbniand An. marshallii s.l. These 2 species are not recognized malaria vectors and thus it is possible thatthe ELISA results are misleading.