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Conservation of the S10-spc-alpha locus within otherwise highly plastic genomes provides phylogenetic insight into the genus Leptospira.

Victoria B, Ahmed A, Zuerner RL, Ahmed N, Bulach DM, Quinteiro J, Hartskeerl RA - PLoS ONE (2008)

Bottom Line: In contrast, PCR analysis of this locus using DNA from saprophytic Leptospira species and species with an intermediate pathogenic capacity generated varied results.Multilocus sequence typing (MLST) of four conserved regions resulted in the construction of well-defined phylogenetic trees that help resolve questions about the interrelationships of pathogenic Leptospira.Based on the results of secY sequence analysis, we found that reliable species identification of pathogenic Leptospira is possible by comparative analysis of a 245 bp region commonly used as a target for diagnostic PCR for leptospirosis.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry and Molecular Biology, Faculty of Biology, University of Santiago de Compostela, Galicia, Spain.

ABSTRACT
S10-spc-alpha is a 17.5 kb cluster of 32 genes encoding ribosomal proteins. This locus has an unusual composition and organization in Leptospira interrogans. We demonstrate the highly conserved nature of this region among diverse Leptospira and show its utility as a phylogenetically informative region. Comparative analyses were performed by PCR using primer sets covering the whole locus. Correctly sized fragments were obtained by PCR from all L. interrogans strains tested for each primer set indicating that this locus is well conserved in this species. Few differences were detected in amplification profiles between different pathogenic species, indicating that the S10-spc-alpha locus is conserved among pathogenic Leptospira. In contrast, PCR analysis of this locus using DNA from saprophytic Leptospira species and species with an intermediate pathogenic capacity generated varied results. Sequence alignment of the S10-spc-alpha locus from two pathogenic species, L. interrogans and L. borgpetersenii, with the corresponding locus from the saprophyte L. biflexa serovar Patoc showed that genetic organization of this locus is well conserved within Leptospira. Multilocus sequence typing (MLST) of four conserved regions resulted in the construction of well-defined phylogenetic trees that help resolve questions about the interrelationships of pathogenic Leptospira. Based on the results of secY sequence analysis, we found that reliable species identification of pathogenic Leptospira is possible by comparative analysis of a 245 bp region commonly used as a target for diagnostic PCR for leptospirosis. Comparative analysis of Leptospira strains revealed that strain H6 previously classified as L. inadai actually belongs to the pathogenic species L. interrogans and that L. meyeri strain ICF phylogenetically co-localized with the pathogenic clusters. These findings demonstrate that the S10-spc-alpha locus is highly conserved throughout the genus and may be more useful in comparing evolution of the genus than loci studied previously.

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Related in: MedlinePlus

Consensus tree based on PCR amplification data.Majority-rule consensus tree elaborated under the parsimony criterion and based on binary data (absence/presence) coded from amplification patterns in the S10-spc-α locus for different Leptospira species. Numbers on nodes are bootstrap support after 100 replicates. Only bootstrap values above or equal to 50% are shown. Species included in the sequence analysis are coded in color. L. biflexa was used as the outgroup. CI = 0.346.
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pone-0002752-g001: Consensus tree based on PCR amplification data.Majority-rule consensus tree elaborated under the parsimony criterion and based on binary data (absence/presence) coded from amplification patterns in the S10-spc-α locus for different Leptospira species. Numbers on nodes are bootstrap support after 100 replicates. Only bootstrap values above or equal to 50% are shown. Species included in the sequence analysis are coded in color. L. biflexa was used as the outgroup. CI = 0.346.

Mentions: The parsimony criterion was used to infer phylogenetic relationships within Leptospira from binary data. The most parsimonious tree generated from these data shows two prominent well-supported clades: 1) a basal clade, with bootstrapping value of 81%, that includes L. fainei and L. inadai, two species with intermediate pathogenic status, and the saprophytic L. meyeri strain Veldrat Semarang 173; and 2) a sister clade, supported with a 68% bootstrap value, that contains pathogenic Leptospira species (Fig. 1). Within the pathogenic clade, relationships among L. alexanderi, L. santarosai, and L. weilii species, are poorly resolved. In contrast, L. interrogans, L. kirschneri, L. borgpetersenii, and L. noguchii are clustered in a well-supported clade where L. kirschneri and L. interrogans appear as closely related siblings.


Conservation of the S10-spc-alpha locus within otherwise highly plastic genomes provides phylogenetic insight into the genus Leptospira.

Victoria B, Ahmed A, Zuerner RL, Ahmed N, Bulach DM, Quinteiro J, Hartskeerl RA - PLoS ONE (2008)

Consensus tree based on PCR amplification data.Majority-rule consensus tree elaborated under the parsimony criterion and based on binary data (absence/presence) coded from amplification patterns in the S10-spc-α locus for different Leptospira species. Numbers on nodes are bootstrap support after 100 replicates. Only bootstrap values above or equal to 50% are shown. Species included in the sequence analysis are coded in color. L. biflexa was used as the outgroup. CI = 0.346.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2481283&req=5

pone-0002752-g001: Consensus tree based on PCR amplification data.Majority-rule consensus tree elaborated under the parsimony criterion and based on binary data (absence/presence) coded from amplification patterns in the S10-spc-α locus for different Leptospira species. Numbers on nodes are bootstrap support after 100 replicates. Only bootstrap values above or equal to 50% are shown. Species included in the sequence analysis are coded in color. L. biflexa was used as the outgroup. CI = 0.346.
Mentions: The parsimony criterion was used to infer phylogenetic relationships within Leptospira from binary data. The most parsimonious tree generated from these data shows two prominent well-supported clades: 1) a basal clade, with bootstrapping value of 81%, that includes L. fainei and L. inadai, two species with intermediate pathogenic status, and the saprophytic L. meyeri strain Veldrat Semarang 173; and 2) a sister clade, supported with a 68% bootstrap value, that contains pathogenic Leptospira species (Fig. 1). Within the pathogenic clade, relationships among L. alexanderi, L. santarosai, and L. weilii species, are poorly resolved. In contrast, L. interrogans, L. kirschneri, L. borgpetersenii, and L. noguchii are clustered in a well-supported clade where L. kirschneri and L. interrogans appear as closely related siblings.

Bottom Line: In contrast, PCR analysis of this locus using DNA from saprophytic Leptospira species and species with an intermediate pathogenic capacity generated varied results.Multilocus sequence typing (MLST) of four conserved regions resulted in the construction of well-defined phylogenetic trees that help resolve questions about the interrelationships of pathogenic Leptospira.Based on the results of secY sequence analysis, we found that reliable species identification of pathogenic Leptospira is possible by comparative analysis of a 245 bp region commonly used as a target for diagnostic PCR for leptospirosis.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry and Molecular Biology, Faculty of Biology, University of Santiago de Compostela, Galicia, Spain.

ABSTRACT
S10-spc-alpha is a 17.5 kb cluster of 32 genes encoding ribosomal proteins. This locus has an unusual composition and organization in Leptospira interrogans. We demonstrate the highly conserved nature of this region among diverse Leptospira and show its utility as a phylogenetically informative region. Comparative analyses were performed by PCR using primer sets covering the whole locus. Correctly sized fragments were obtained by PCR from all L. interrogans strains tested for each primer set indicating that this locus is well conserved in this species. Few differences were detected in amplification profiles between different pathogenic species, indicating that the S10-spc-alpha locus is conserved among pathogenic Leptospira. In contrast, PCR analysis of this locus using DNA from saprophytic Leptospira species and species with an intermediate pathogenic capacity generated varied results. Sequence alignment of the S10-spc-alpha locus from two pathogenic species, L. interrogans and L. borgpetersenii, with the corresponding locus from the saprophyte L. biflexa serovar Patoc showed that genetic organization of this locus is well conserved within Leptospira. Multilocus sequence typing (MLST) of four conserved regions resulted in the construction of well-defined phylogenetic trees that help resolve questions about the interrelationships of pathogenic Leptospira. Based on the results of secY sequence analysis, we found that reliable species identification of pathogenic Leptospira is possible by comparative analysis of a 245 bp region commonly used as a target for diagnostic PCR for leptospirosis. Comparative analysis of Leptospira strains revealed that strain H6 previously classified as L. inadai actually belongs to the pathogenic species L. interrogans and that L. meyeri strain ICF phylogenetically co-localized with the pathogenic clusters. These findings demonstrate that the S10-spc-alpha locus is highly conserved throughout the genus and may be more useful in comparing evolution of the genus than loci studied previously.

Show MeSH
Related in: MedlinePlus