Comparison of Two Multilocus Sequence Based Genotyping Schemes for Leptospira Species
Author(s)
Type
Journal Article
Abstract
Background: Several sequence based genotyping schemes have been developed for Leptospira spp. The objective of this
study was to genotype a collection of clinical and reference isolates using the two most commonly used schemes and
compare and contrast the results.
Methods and Findings: A total of 48 isolates consisting of L. interrogans (n = 40) and L. kirschneri (n = 8) were typed by the 7
locus MLST scheme described by Thaipadungpanit et al., and the 6 locus genotyping scheme described by Ahmed et al.,
(termed 7L and 6L, respectively). Two L. interrogans isolates were not typed using 6L because of a deletion of three
nucleotides in lipL32. The remaining 46 isolates were resolved into 21 sequence types (STs) by 7L, and 30 genotypes by 6L.
Overall nucleotide diversity (based on concatenated sequence) was 3.6% and 2.3% for 7L and 6L, respectively. The D value
(discriminatory ability) of 7L and 6L were comparable, i.e. 92.0 (95% CI 87.5–96.5) vs. 93.5 (95% CI 88.6–98.4). The dN/dS
ratios calculated for each locus indicated that none were under positive selection. Neighbor joining trees were
reconstructed based on the concatenated sequences for each scheme. Both trees showed two distinct groups
corresponding to L. interrogans and L. kirschneri, and both identified two clones containing 10 and 7 clinical isolates,
respectively. There were six instances in which 6L split single STs as defined by 7L into closely related clusters. We noted two
discrepancies between the trees in which the genetic relatedness between two pairs of strains were more closely related by
7L than by 6L.
Conclusions: This genetic analysis indicates that the two schemes are comparable. We discuss their practical advantages
and disadvantages.
study was to genotype a collection of clinical and reference isolates using the two most commonly used schemes and
compare and contrast the results.
Methods and Findings: A total of 48 isolates consisting of L. interrogans (n = 40) and L. kirschneri (n = 8) were typed by the 7
locus MLST scheme described by Thaipadungpanit et al., and the 6 locus genotyping scheme described by Ahmed et al.,
(termed 7L and 6L, respectively). Two L. interrogans isolates were not typed using 6L because of a deletion of three
nucleotides in lipL32. The remaining 46 isolates were resolved into 21 sequence types (STs) by 7L, and 30 genotypes by 6L.
Overall nucleotide diversity (based on concatenated sequence) was 3.6% and 2.3% for 7L and 6L, respectively. The D value
(discriminatory ability) of 7L and 6L were comparable, i.e. 92.0 (95% CI 87.5–96.5) vs. 93.5 (95% CI 88.6–98.4). The dN/dS
ratios calculated for each locus indicated that none were under positive selection. Neighbor joining trees were
reconstructed based on the concatenated sequences for each scheme. Both trees showed two distinct groups
corresponding to L. interrogans and L. kirschneri, and both identified two clones containing 10 and 7 clinical isolates,
respectively. There were six instances in which 6L split single STs as defined by 7L into closely related clusters. We noted two
discrepancies between the trees in which the genetic relatedness between two pairs of strains were more closely related by
7L than by 6L.
Conclusions: This genetic analysis indicates that the two schemes are comparable. We discuss their practical advantages
and disadvantages.
Date Issued
2011-11-01
Date Acceptance
2011-09-13
Citation
PLOS Neglected Tropical Diseases, 2011, 5 (11)
ISSN
1935-2735
Publisher
Public Library of Science
Journal / Book Title
PLOS Neglected Tropical Diseases
Volume
5
Issue
11
Copyright Statement
© 2011 Ahmed et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits
unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
License URL
Subjects
Science & Technology
Life Sciences & Biomedicine
Infectious Diseases
Parasitology
Tropical Medicine
PARASITOLOGY
TROPICAL MEDICINE
FRAGMENT-LENGTH-POLYMORPHISMS
ACUTE FEBRILE ILLNESS
REAL-TIME PCR
FAMILY LEPTOSPIRACEAE
VARIABLE-NUMBER
SEROVAR HARDJO
SRI-LANKA
IDENTIFICATION
INTERROGANS
DNA
Publication Status
Published
Article Number
e1374