GenBank accession numbers are provided as additional file 1 Tables S1 and S2. Table 4 Leptospira www.selleckchem.com/products/BAY-73-4506.html clusters identified using lfb1 sequence polymorphism. Clusters Serovars Reference Strains Collection isolates Clinical Samples (number of amplicons) % of PCR-diagnosed human cases (January 2008-February 2010) L.interrogans 1 Copenhageni/Pyrogenes 5 isolates Human GSK1210151A solubility dmso (60) and deer (2) 68.2% L.interrogans 2 Autumnalis/Australis/Lai no isolate Human (6) 6.8% L.interrogans 3 Bataviae no isolate Human (3) 3.4% L.interrogans 4 Canicola/Pomona 5 isolates Human

(2) and deer (3) 2.3% L.interrogans 5 Unidentified serovar 5 isolates Human (10) 11.4% L.borgpetersenii 1 Castellonis/Sejroe 4 isolates Human (7) and deer (1) 7.9% L.borgpetersenii 2 Hardjo-bovis 1 isolate Deer (6) 0% We also evaluated if the direct

sequencing of the secY diagnostic product [9] could confirm the existence of the different clusters identified using lfb1 polymorphism (Figure 2). The 202 bp PCR product could successfully be amplified and sequenced from DNA extracted from all isolates. Using DNA from clinical specimens, samples from both lfb1-deduced clusters of L. borgpetersenii were successfully amplified and sequenced, but only samples from 3 out of the 5 lfb1-deduced clusters of L. interrogans could be amplified (clusters L. interrogans 1, 4 and 5). However, samples from the two remaining clusters (clusters L. interrogans 2 and 3) were scarce (see Table 4) and had low Leptospira concentrations (see Table 2). secY products using DNA Epothilone B (EPO906, Patupilone) from these clinical specimens could not be generated, even using combinations of primers used for BIX 1294 mouse the MLST study [18] and for diagnosis [9]. However, the phylogeny deduced from a 174 bp alignment of the diagnostic secY product confirmed the clusters identified by both the MLST and lfb1 typing schemes. Strains from cluster L. interrogans 5 had sequences 100% identical to L. interrogans Hardjo-prajitno (strain Hardjoprajitno) and to L. meyeri serovar Perameles strain Bandicoot, a strain recently re-assigned to the species L. interrogans [25]. GenBank

accession numbers of the sequences generated and used in this study are provided as additional file 1 Tables S1 and S2. Figure 2 secY -derived phylogeny of New Caledonian isolates, clinical specimens and reference strains based on a 174 bp sequence polymorphism. Blue legends indicate reference strains, red legends indicate the putative unknown serovar. GenBank accession numbers are provided as additional file 1 Tables S1 and S2. MLST-deduced phylogeny DNA sequences retrieved from databases or sequenced from products successfully amplified were concatenated and allowed drawing a phylogeny of the New Caledonian isolates, together with reference strains (Figure 3). GenBank accession numbers of the sequences generated and used in this study are provided as additional file 1 Tables S1 and S2.