title: Analysis of 16S rDNA sequences from pathogenic Leptospira serovars and use of single nucleotide polymorphisms for rapid speciation by D-HPLC.
abstract: Leptospira have a worldwide distribution and include important zoonotic pathogens yet diagnosis and differentiation still tend to rely on traditional bacteriological and serological approaches. In this study a 1.3 kb fragment of the rrs gene (16S rDNA) was sequenced from a panel of 22 control strains, representing serovars within the pathogenic species Leptospira interrogans, Leptospiraborgpetersenii, and Leptospirakirschneri, to identify single nucleotide polymorphisms (SNPs). These were identified in the 5' variable region of the 16S sequence and a 181 bp PCR fragment encompassing this region was used for speciation by Denaturing High Performance Liquid Chromatography (D-HPLC). This method was applied to eleven additional species, representing pathogenic, non-pathogenic and intermediate species and was demonstrated to rapidly differentiate all but 2 of the non-pathogenic Leptospira species. The method was applied successfully to infected tissues from field samples proving its value for diagnosing leptospiral infections found in animals in the UK.
authors.affiliation: Department of Food and Environmental Safety, Veterinary Laboratories Agency (Weybridge), Woodham Lane, New Haw, Addlestone, Surrey KT15 3NB, UK. j.fenner@vla.defra.gsi.gov.uk
authors.forename: J S
authors.initials: JS
authors.lastname: Fenner
authors.forename: M F
authors.initials: MF
authors.lastname: Anjum
authors.forename: L P
authors.initials: LP
authors.lastname: Randall
authors.forename: G C
authors.initials: GC
authors.lastname: Pritchard
authors.forename: G
authors.initials: G
authors.lastname: Wu
authors.forename: J
authors.initials: J
authors.lastname: Errington
authors.forename: C G
authors.initials: CG
authors.lastname: Dalley
authors.forename: M J
authors.initials: MJ
authors.lastname: Woodward
mesh_terms: D001483:Base Sequence; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D015994:Incidence; D007919:Leptospira / Q000145:classification* / Q000235:genetics / Q000472:pathogenicity*; D008969:Molecular Sequence Data; D010802:Phylogeny; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*; D012329:RNA, Bacterial / Q000235:genetics*; D012336:RNA, Ribosomal, 16S / Q000235:genetics*; D013045:Species Specificity
chemical_list: D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S