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Profiling of Leptospira interrogans, L. santarosai, L. meyeri and L. borgpetersenii by SE-AFLP, PFGE and susceptibility testing—a continuous attempt at species and serovar differentiation

Leptospirosis is a widespread systemic zoonosis, considered as reemerging in certain developing countries. Although the cross agglutinin absorption test is still considered the standard method for Leptospira identification, it presents several disadvantages. The aim of this study was to characterize...

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Autores principales: Moreno, Luisa Z, Miraglia, Fabiana, Lilenbaum, Walter, Neto, José SF, Freitas, Julio C, Morais, Zenaide M, Hartskeerl, Rudy A, da Costa, Barbara LP, Vasconcellos, Silvio A, Moreno, Andrea M
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4820670/
https://www.ncbi.nlm.nih.gov/pubmed/26956446
http://dx.doi.org/10.1038/emi.2016.16
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author Moreno, Luisa Z
Miraglia, Fabiana
Lilenbaum, Walter
Neto, José SF
Freitas, Julio C
Morais, Zenaide M
Hartskeerl, Rudy A
da Costa, Barbara LP
Vasconcellos, Silvio A
Moreno, Andrea M
author_facet Moreno, Luisa Z
Miraglia, Fabiana
Lilenbaum, Walter
Neto, José SF
Freitas, Julio C
Morais, Zenaide M
Hartskeerl, Rudy A
da Costa, Barbara LP
Vasconcellos, Silvio A
Moreno, Andrea M
author_sort Moreno, Luisa Z
collection PubMed
description Leptospirosis is a widespread systemic zoonosis, considered as reemerging in certain developing countries. Although the cross agglutinin absorption test is still considered the standard method for Leptospira identification, it presents several disadvantages. The aim of this study was to characterize Leptospira spp. isolated from various hosts by genotyping and broth microdilution susceptibility testing in an attempt to differentiate Leptospira species, serogroups and serovars. Forty-seven isolates were studied. They were previously serotyped, and species confirmation was performed by 16S rRNA sequencing. Single-enzyme amplified fragment length polymorphism (SE-AFLP) and pulsed-field gel electrophoresis (PFGE) analysis enabled the distinction of L. interrogans from L. santarosai, L. meyeri and L. borgpetersenii in two main clusters. Among L. interrogans, it was possible to differentiate into two new clusters the serogroup Icterohaemorrhagiae from the serogroups Canicola and Pomona. L. santarosai isolates presented higher genetic variation than the other species in both techniques. Interestingly, the minimum inhibitory concentration (MIC) cluster analysis also provided Leptospira serogroup differentiation. Further studies are necessary regarding serovar Bananal isolates, as they presented the highest MIC values for most of the antimicrobials tested. All studied techniques successfully distinguished Leptospira species and serogroups. Despite being library-dependent methods, these approaches are less labor intensive and more economically viable, particularly SE-AFLP, and can be implemented in most reference laboratories worldwide to enable faster Leptospira typing.
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spelling pubmed-48206702016-04-17 Profiling of Leptospira interrogans, L. santarosai, L. meyeri and L. borgpetersenii by SE-AFLP, PFGE and susceptibility testing—a continuous attempt at species and serovar differentiation Moreno, Luisa Z Miraglia, Fabiana Lilenbaum, Walter Neto, José SF Freitas, Julio C Morais, Zenaide M Hartskeerl, Rudy A da Costa, Barbara LP Vasconcellos, Silvio A Moreno, Andrea M Emerg Microbes Infect Original Article Leptospirosis is a widespread systemic zoonosis, considered as reemerging in certain developing countries. Although the cross agglutinin absorption test is still considered the standard method for Leptospira identification, it presents several disadvantages. The aim of this study was to characterize Leptospira spp. isolated from various hosts by genotyping and broth microdilution susceptibility testing in an attempt to differentiate Leptospira species, serogroups and serovars. Forty-seven isolates were studied. They were previously serotyped, and species confirmation was performed by 16S rRNA sequencing. Single-enzyme amplified fragment length polymorphism (SE-AFLP) and pulsed-field gel electrophoresis (PFGE) analysis enabled the distinction of L. interrogans from L. santarosai, L. meyeri and L. borgpetersenii in two main clusters. Among L. interrogans, it was possible to differentiate into two new clusters the serogroup Icterohaemorrhagiae from the serogroups Canicola and Pomona. L. santarosai isolates presented higher genetic variation than the other species in both techniques. Interestingly, the minimum inhibitory concentration (MIC) cluster analysis also provided Leptospira serogroup differentiation. Further studies are necessary regarding serovar Bananal isolates, as they presented the highest MIC values for most of the antimicrobials tested. All studied techniques successfully distinguished Leptospira species and serogroups. Despite being library-dependent methods, these approaches are less labor intensive and more economically viable, particularly SE-AFLP, and can be implemented in most reference laboratories worldwide to enable faster Leptospira typing. Nature Publishing Group 2016-03 2016-03-09 /pmc/articles/PMC4820670/ /pubmed/26956446 http://dx.doi.org/10.1038/emi.2016.16 Text en Copyright © 2016 Shanghai Shangyixun Cultural Communication Co., Ltd http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
spellingShingle Original Article
Moreno, Luisa Z
Miraglia, Fabiana
Lilenbaum, Walter
Neto, José SF
Freitas, Julio C
Morais, Zenaide M
Hartskeerl, Rudy A
da Costa, Barbara LP
Vasconcellos, Silvio A
Moreno, Andrea M
Profiling of Leptospira interrogans, L. santarosai, L. meyeri and L. borgpetersenii by SE-AFLP, PFGE and susceptibility testing—a continuous attempt at species and serovar differentiation
title Profiling of Leptospira interrogans, L. santarosai, L. meyeri and L. borgpetersenii by SE-AFLP, PFGE and susceptibility testing—a continuous attempt at species and serovar differentiation
title_full Profiling of Leptospira interrogans, L. santarosai, L. meyeri and L. borgpetersenii by SE-AFLP, PFGE and susceptibility testing—a continuous attempt at species and serovar differentiation
title_fullStr Profiling of Leptospira interrogans, L. santarosai, L. meyeri and L. borgpetersenii by SE-AFLP, PFGE and susceptibility testing—a continuous attempt at species and serovar differentiation
title_full_unstemmed Profiling of Leptospira interrogans, L. santarosai, L. meyeri and L. borgpetersenii by SE-AFLP, PFGE and susceptibility testing—a continuous attempt at species and serovar differentiation
title_short Profiling of Leptospira interrogans, L. santarosai, L. meyeri and L. borgpetersenii by SE-AFLP, PFGE and susceptibility testing—a continuous attempt at species and serovar differentiation
title_sort profiling of leptospira interrogans, l. santarosai, l. meyeri and l. borgpetersenii by se-aflp, pfge and susceptibility testing—a continuous attempt at species and serovar differentiation
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4820670/
https://www.ncbi.nlm.nih.gov/pubmed/26956446
http://dx.doi.org/10.1038/emi.2016.16
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