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Development of a comparative genomic fingerprinting assay for rapid and high resolution genotyping of Arcobacter butzleri
BACKGROUND: Molecular typing methods are critical for epidemiological investigations, facilitating disease outbreak detection and source identification. Study of the epidemiology of the emerging human pathogen Arcobacter butzleri is currently hampered by the lack of a subtyping method that is easily...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4424573/ https://www.ncbi.nlm.nih.gov/pubmed/25947176 http://dx.doi.org/10.1186/s12866-015-0426-4 |
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author | Webb, Andrew L Kruczkiewicz, Peter Selinger, L Brent Inglis, G Douglas Taboada, Eduardo N |
author_facet | Webb, Andrew L Kruczkiewicz, Peter Selinger, L Brent Inglis, G Douglas Taboada, Eduardo N |
author_sort | Webb, Andrew L |
collection | PubMed |
description | BACKGROUND: Molecular typing methods are critical for epidemiological investigations, facilitating disease outbreak detection and source identification. Study of the epidemiology of the emerging human pathogen Arcobacter butzleri is currently hampered by the lack of a subtyping method that is easily deployable in the context of routine epidemiological surveillance. In this study we describe a comparative genomic fingerprinting (CGF) method for high-resolution and high-throughput subtyping of A. butzleri. Comparative analysis of the genome sequences of eleven A. butzleri strains, including eight strains newly sequenced as part of this project, was employed to identify accessory genes suitable for generating unique genetic fingerprints for high-resolution subtyping based on gene presence or absence within a strain. RESULTS: A set of eighty-three accessory genes was used to examine the population structure of a dataset comprised of isolates from various sources, including human and non-human animals, sewage, and river water (n=156). A streamlined assay (CGF(40)) based on a subset of 40 genes was subsequently developed through marker optimization. High levels of profile diversity (121 distinct profiles) were observed among the 156 isolates in the dataset, and a high Simpson’s Index of Diversity (ID) observed (ID > 0.969) indicate that the CGF(40) assay possesses high discriminatory power. At the same time, our observation that 115 isolates in this dataset could be assigned to 29 clades with a profile similarity of 90% or greater indicates that the method can be used to identify clades comprised of genetically similar isolates. CONCLUSIONS: The CGF(40) assay described herein combines high resolution and repeatability with high throughput for the rapid characterization of A. butzleri strains. This assay will facilitate the study of the population structure and epidemiology of A. butzleri. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12866-015-0426-4) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-4424573 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-44245732015-05-09 Development of a comparative genomic fingerprinting assay for rapid and high resolution genotyping of Arcobacter butzleri Webb, Andrew L Kruczkiewicz, Peter Selinger, L Brent Inglis, G Douglas Taboada, Eduardo N BMC Microbiol Methodology Article BACKGROUND: Molecular typing methods are critical for epidemiological investigations, facilitating disease outbreak detection and source identification. Study of the epidemiology of the emerging human pathogen Arcobacter butzleri is currently hampered by the lack of a subtyping method that is easily deployable in the context of routine epidemiological surveillance. In this study we describe a comparative genomic fingerprinting (CGF) method for high-resolution and high-throughput subtyping of A. butzleri. Comparative analysis of the genome sequences of eleven A. butzleri strains, including eight strains newly sequenced as part of this project, was employed to identify accessory genes suitable for generating unique genetic fingerprints for high-resolution subtyping based on gene presence or absence within a strain. RESULTS: A set of eighty-three accessory genes was used to examine the population structure of a dataset comprised of isolates from various sources, including human and non-human animals, sewage, and river water (n=156). A streamlined assay (CGF(40)) based on a subset of 40 genes was subsequently developed through marker optimization. High levels of profile diversity (121 distinct profiles) were observed among the 156 isolates in the dataset, and a high Simpson’s Index of Diversity (ID) observed (ID > 0.969) indicate that the CGF(40) assay possesses high discriminatory power. At the same time, our observation that 115 isolates in this dataset could be assigned to 29 clades with a profile similarity of 90% or greater indicates that the method can be used to identify clades comprised of genetically similar isolates. CONCLUSIONS: The CGF(40) assay described herein combines high resolution and repeatability with high throughput for the rapid characterization of A. butzleri strains. This assay will facilitate the study of the population structure and epidemiology of A. butzleri. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12866-015-0426-4) contains supplementary material, which is available to authorized users. BioMed Central 2015-05-07 /pmc/articles/PMC4424573/ /pubmed/25947176 http://dx.doi.org/10.1186/s12866-015-0426-4 Text en © Webb et al.; licensee BioMed Central. 2015 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Methodology Article Webb, Andrew L Kruczkiewicz, Peter Selinger, L Brent Inglis, G Douglas Taboada, Eduardo N Development of a comparative genomic fingerprinting assay for rapid and high resolution genotyping of Arcobacter butzleri |
title | Development of a comparative genomic fingerprinting assay for rapid and high resolution genotyping of Arcobacter butzleri |
title_full | Development of a comparative genomic fingerprinting assay for rapid and high resolution genotyping of Arcobacter butzleri |
title_fullStr | Development of a comparative genomic fingerprinting assay for rapid and high resolution genotyping of Arcobacter butzleri |
title_full_unstemmed | Development of a comparative genomic fingerprinting assay for rapid and high resolution genotyping of Arcobacter butzleri |
title_short | Development of a comparative genomic fingerprinting assay for rapid and high resolution genotyping of Arcobacter butzleri |
title_sort | development of a comparative genomic fingerprinting assay for rapid and high resolution genotyping of arcobacter butzleri |
topic | Methodology Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4424573/ https://www.ncbi.nlm.nih.gov/pubmed/25947176 http://dx.doi.org/10.1186/s12866-015-0426-4 |
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