title: Rapid Leptospira identification by direct sequencing of the diagnostic PCR products in New Caledonia.
abstract: BACKGROUND
  Most of the current knowledge of leptospirosis epidemiology originates from serological results obtained with the reference Microscopic Agglutination Test (MAT). However, inconsistencies and weaknesses of this diagnostic technique are evident. A growing use of PCR has improved the early diagnosis of leptospirosis but a drawback is that it cannot provide information on the infecting Leptospira strain which provides important epidemiologic data. Our work is aimed at evaluating if the sequence polymorphism of diagnostic PCR products could be used to identify the infecting Leptospira strains in the New Caledonian environment.
  
  
  RESULTS
  Both the lfb1 and secY diagnostic PCR products displayed a sequence polymorphism that could prove useful in presumptively identifying the infecting leptospire. Using both this polymorphism and MLST results with New Caledonian isolates and clinical samples, we confirmed the epidemiological relevance of the sequence-based identification of Leptospira strains. Additionally, we identified one cluster of L. interrogans that contained no reference strain and one cluster of L. borgpetersenii found only in the introduced Rusa deer Cervus timorensis russa that is its probable reservoir.
  
  
  CONCLUSIONS
  The sequence polymorphism of diagnostic PCR products proved useful in presumptively identifying the infecting Leptospira strains. This could contribute to a better understanding of leptospirosis epidemiology by providing epidemiological information that cannot be directly attained from the use of PCR as an early diagnostic test for leptospirosis.
authors.affiliation: Institut Pasteur de Nouvelle-Calédonie, Réseau International des Instituts Pasteur, Laboratoire de Recherche en Bactériologie, BP61, 98845 Nouméa cedex, New Caledonia.
authors.forename: Julie
authors.initials: J
authors.lastname: Perez
authors.forename: Cyrille
authors.initials: C
authors.lastname: Goarant
mesh_terms: D017931:DNA Primers / Q000235:genetics; D006801:Humans; D007919:Leptospira / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D007922:Leptospirosis / Q000175:diagnosis* / Q000453:epidemiology / Q000382:microbiology*; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D008969:Molecular Sequence Data; D009510:New Caledonia / Q000453:epidemiology; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA / Q000379:methods
chemical_list: D017931:DNA Primers