To determine the past prevalence of HIV-1 and HIV-2 in West Africa, we tested 440 serum samples collected in 1980 from people living in rural areas of Guinea Bissau. The sera were screened for antibodies by enzyme immunoassay (EIA) using purified simian immunodeficiency virus (SIV) isolated from rhesus macaque monkeys (SIVmac) and then confirmed by immunoblot and immunofluorescence assays for antibodies to SIV, HIV-2, and HIV-1. Five of the serum samples were repeatedly reactive by all assays to both SIV and HIV-2. None were positive for antibodies to HIV-1 by immunofluorescence, immunoblot or radio-immunoprecipitation (RIP) assays, with the exception of one HIV-2-seropositive samples which reacted with HIV-1 glycoproteins by RIPA. Thus, in 1980, at least 1.4% (six out of 440) of a random sample of people in rural Guinea Bissau had been exposed to a virus highly related to HIV-2 and SIV. One of these subjects may have been dually infected with HIV-1 and HIV-2.