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Use of a New High Resolution Melting Method for Genotyping Pathogenic Leptospira spp
- Source :
- PLoS ONE, PLoS ONE, 2015, 10 (7), pp.e0127430. ⟨10.1371/journal.pone.0127430⟩, PLoS ONE, Public Library of Science, 2015, 10 (7), pp.e0127430. ⟨10.1371/journal.pone.0127430⟩, PLoS ONE, Vol 10, Iss 7, p e0127430 (2015)
- Publication Year :
- 2015
- Publisher :
- HAL CCSD, 2015.
-
Abstract
- Background Leptospirosis is a worldwide zoonosis that is endemic in tropical areas, such as Reunion Island. The species Leptospira interrogans is the primary agent in human infections, but other pathogenic species, such as L. kirschner and L. borgpetersenii, are also associated with human leptospirosis. Methods and Findings In this study, a melting curve analysis of the products that were amplified with the primer pairs lfb1 F/R and G1/G2 facilitated an accurate species classification of Leptospira reference strains. Next, we combined an unsupervised high resolution melting (HRM) method with a new statistical approach using primers to amplify a two variable-number tandem-repeat (VNTR) for typing at the subspecies level. The HRM analysis, which was performed with ScreenClust Software, enabled the identification of genotypes at the serovar level with high resolution power (Hunter-Gaston index 0.984). This method was also applied to Leptospira DNA from blood samples that were obtained from Reunion Island after 1998. We were able to identify a unique genotype that is identical to that of the L. interrogans serovars Copenhageni and Icterohaemorrhagiae, suggesting that this genotype is the major cause of leptospirosis on Reunion Island. Conclusions Our simple, rapid, and robust genotyping method enables the identification of Leptospira strains at the species and subspecies levels and supports the direct genotyping of Leptospira in biological samples without requiring cultures.
- Subjects :
- Serotype
DNA, Bacterial
Genotype
Genotyping Techniques
[SDV]Life Sciences [q-bio]
lcsh:Medicine
Minisatellite Repeats
Nucleic Acid Denaturation
Real-Time Polymerase Chain Reaction
Melting curve analysis
High Resolution Melt
Species Specificity
Leptospira
medicine
Cluster Analysis
Humans
lcsh:Science
Genotyping
Genetics
Multidisciplinary
biology
lcsh:R
biology.organism_classification
medicine.disease
Leptospirosis
Virology
[SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology
lcsh:Q
Leptospira interrogans
Software
Research Article
Subjects
Details
- Language :
- English
- ISSN :
- 19326203
- Database :
- OpenAIRE
- Journal :
- PLoS ONE, PLoS ONE, 2015, 10 (7), pp.e0127430. ⟨10.1371/journal.pone.0127430⟩, PLoS ONE, Public Library of Science, 2015, 10 (7), pp.e0127430. ⟨10.1371/journal.pone.0127430⟩, PLoS ONE, Vol 10, Iss 7, p e0127430 (2015)
- Accession number :
- edsair.doi.dedup.....29abe719c6db2bc09ff09cab78bfda02