Isolations of Mokola virus (MOKV) are rare, but in South Africa and Zimbabwe this genotype 3 lyssavirus variant has been occasionally found in domestic mammals (cats and a dog) with a total of 17 virus isolates (South Africa 10, Zimbabwe 7) having been recovered during the past 30 years. We report the identification of a MOKV isolate involved in a human contact in Grahamstown (Eastern Cape, South Africa) and a genetic comparison with previously characterized isolates. This reported MOKV case was in a previously immunized cat. While the continual recovery of MOKV isolates in domestic cats is speculative of the existence of a reservoir host species among bats or rodents, the lack of protection with currently used vaccines is discussed and the need for biologicals with a wider spectrum of protection against this lyssavirus variant is highlighted.