Abstract We have amplified Mycobacterium leprae DNA from the skeleton of an adult human female exhibiting signs of lepromatous leprosy (LL). The remains were excavated from the site of Devkesken 6 on the Ustyurt plateau of Uzbekistan and date to between the 1st and 4th centuries AD. Recovered DNA was fragmented but of sufficient quality and quantity to allow a series of biomolecular genotyping methods to be applied. These methods included variable nucleotide tandem repeat (VNTR) typing of two microsatellite and one minisatellite regions and also single-nucleotide polymorphism (SNP) typing for nine informative loci. Genotyping showed that the causative strain of M. leprae exhibited a SNP-type 3 profile, characteristic of cases associated geographically with Europe and North Africa. Further SNP sub-typing was performed and the data obtained from the Uzbek leper was compared with the same loci amplified from a case of LL recovered from Blackfriars, Ipswich, UK dating to between the 13th and 16th centuries AD. Unique group 3 subtypes were found in both the Uzbek case and Ipswich 1914. These appear to be ancestral to recent type 3 strains. Mycolic acid analysis confirmed the presence of M. leprae in the Uzbek samples. Phylogenetically informative SNPs and other polymorphic loci will contribute to the study of human migrations, as well as the origin and spread of leprosy.