title: Genotype of a historic strain of Mycobacterium tuberculosis.
abstract: The use of ancient DNA in paleopathological studies of tuberculosis has largely been restricted to confirmation of disease identifications made by skeletal analysis; few attempts at obtaining genotype data from archaeological samples have been made because of the need to perform different PCRs for each genetic locus being studied in an ancient DNA extract. We used a next generation sequencing approach involving hybridization capture directed at specific polymorphic regions of the Mycobacterium tuberculosis genome to identify a detailed genotype for a historic strain of M. tuberculosis from an individual buried in the 19th century St. George's Crypt, Leeds, West Yorkshire, England. We obtained 664,500 sequencing by oligonucleotide ligation and detection (SOLiD) reads that mapped to the targeted regions of the M. tuberculosis genome; the coverage included 218 of 247 SNPs, 10 of 11 insertion/deletion regions, and the repeat elements IS1081 and IS6110. The accuracy of the SOLiD data was checked by conventional PCRs directed at 11 SNPs and two insertion/deletions. The data placed the historic strain of M. tuberculosis in a group that is uncommon today, but it is known to have been present in North America in the early 20th century. Our results show the use of hybridization capture followed by next generation sequencing as a means of obtaining detailed genotypes of ancient varieties of M. tuberculosis, potentially enabling meaningful comparisons between strains from different geographic locations and different periods in the past.
authors.affiliation: Manchester Institute of Biotechnology, Faculty of Life Sciences, University of Manchester, Manchester M1 7DN, United Kingdom.
authors.forename: Abigail S
authors.initials: AS
authors.lastname: Bouwman
authors.forename: Sandra L
authors.initials: SL
authors.lastname: Kennedy
authors.forename: Romy
authors.initials: R
authors.lastname: Müller
authors.forename: Richard H
authors.initials: RH
authors.lastname: Stephens
authors.forename: Malin
authors.initials: M
authors.lastname: Holst
authors.forename: Anwen C
authors.initials: AC
authors.lastname: Caffell
authors.forename: Charlotte A
authors.initials: CA
authors.lastname: Roberts
authors.forename: Terence A
authors.initials: TA
authors.lastname: Brown
mesh_terms: D015373:Bacterial Typing Techniques; D001483:Base Sequence; D004739:England; D005260:Female; D016680:Genome, Bacterial / Q000235:genetics; D005838:Genotype; D006801:Humans; D008969:Molecular Sequence Data; D009169:Mycobacterium tuberculosis / Q000235:genetics*; D009841:Oligonucleotides / Q000235:genetics; D010163:Paleontology; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
chemical_list: D009841:Oligonucleotides