A study of host preference in tsetse flies using a modified heteroduplex PCR-based method is described. Domestic and wild animal blood samples were collected to extract the corresponding reference DNAs. In Campo (south Cameroon), tsetse flies (mainly Glossina palpalis palpalis) were trapped and 41 bloodmeals were collected. All reference DNAs and 37 bloodmeal DNAs (90.7%) were successfully amplified and hybridised. Twelve bloodmeals (32.4%) were of human origin, 13 (35.4%) were from Sitatunga (Tragelaphus spekei) (an antelope) while 12 (32.4%) were not identified using our set of reference DNAs. The results confirmed the occurrence of frequent contacts between wild animals and this population of tsetse flies.