Use of a New High Resolution Melting Method for Genotyping Pathogenic Leptospira spp
| Autor: | Amélie Desvars, Alain Michault, Florence Naze, Pascale Bourhy, Mathieu Picardeau |
|---|---|
| Přispěvatelé: | Laboratoire de Bactériologie-Parasitologie-Virologie-Hygiène, Groupe Hospitalier Sud Réunion (GHSR), Centre Collaborateur FAO/OMS pour l'épidémiologie de la leptospirose, Institut Pasteur [Paris] (IP), Centre National de Référence de la Leptospirose - National Reference Center Leptospirosis (CNR), Biologie des Spirochètes / Biology of Spirochetes, The authors have no support or funding to report., Conceived and designed the experiments: AM MP PB. Performed the experiments: FN AD. Analyzed the data: AD MP AM PB. Contributed reagents/materials/analysis tools: FN PB. Wrote the paper: FN AD MP PB AM., Institut Pasteur [Paris] |
| Jazyk: | angličtina |
| Rok vydání: | 2015 |
| Předmět: |
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 |
| Zdroj: | 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) |
| ISSN: | 1932-6203 |
| DOI: | 10.1371/journal.pone.0127430⟩ |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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