Tick-borne relapsing fever caused by the spirochaete Borrelia duttonii is a common cause of serious illness in central Tanzania. Screening of Ornithodoros sp ticks from infested houses for the presence of B duttonii had detected a previously unidentified species of Borrelia. We investigated whether this species infected the human population in a central Tanzanian village, by use of blood slide examination and PCR. PCR was twice as sensitive in detection of infections, showing Borrelia sp in six (11%) of 54 children with fever, and in 13 (4%) of 307 otherwise healthy children. Genotyping Borrelia from 17 infections identified Borrelia duttonii and an unnamed species. Our findings show that the newly discovered species is a causal agent of tick-borne relapsing fever.