Cargando…

Fast DNA Serotyping and Antimicrobial Resistance Gene Determination of Salmonella enterica with an Oligonucleotide Microarray-Based Assay

Salmonellosis caused by Salmonella (S.) belongs to the most prevalent food-borne zoonotic diseases throughout the world. Therefore, serotype identification for all culture-confirmed cases of Salmonella infection is important for epidemiological purposes. As a standard, the traditional culture method...

Descripción completa

Detalles Bibliográficos
Autores principales: Braun, Sascha D., Ziegler, Albrecht, Methner, Ulrich, Slickers, Peter, Keiling, Silke, Monecke, Stefan, Ehricht, Ralf
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3464306/
https://www.ncbi.nlm.nih.gov/pubmed/23056321
http://dx.doi.org/10.1371/journal.pone.0046489
_version_ 1782245406783045632
author Braun, Sascha D.
Ziegler, Albrecht
Methner, Ulrich
Slickers, Peter
Keiling, Silke
Monecke, Stefan
Ehricht, Ralf
author_facet Braun, Sascha D.
Ziegler, Albrecht
Methner, Ulrich
Slickers, Peter
Keiling, Silke
Monecke, Stefan
Ehricht, Ralf
author_sort Braun, Sascha D.
collection PubMed
description Salmonellosis caused by Salmonella (S.) belongs to the most prevalent food-borne zoonotic diseases throughout the world. Therefore, serotype identification for all culture-confirmed cases of Salmonella infection is important for epidemiological purposes. As a standard, the traditional culture method (ISO 6579:2002) is used to identify Salmonella. Classical serotyping takes 4–5 days to be completed, it is labor-intensive, expensive and more than 250 non-standardized sera are necessary to characterize more than 2,500 Salmonella serovars currently known. These technical difficulties could be overcome with modern molecular methods. We developed a microarray based serogenotyping assay for the most prevalent Salmonella serovars in Europe and North America. The current assay version could theoretically discriminate 28 O-antigens and 86 H-antigens. Additionally, we included 77 targets analyzing antimicrobial resistance genes. The Salmonella assay was evaluated with a set of 168 reference strains representing 132 serovars previously serotyped by conventional agglutination through various reference centers. 117 of 132 (81%) tested serovars showed an unique microarray pattern. 15 of 132 serovars generated a pattern which was shared by multiple serovars (e.g., S. ser. Enteritidis and S. ser. Nitra). These shared patterns mainly resulted from the high similarity of the genotypes of serogroup A and D1. Using patterns of the known reference strains, a database was build which represents the basis of a new PatternMatch software that can serotype unknown Salmonella isolates automatically. After assay verification, the Salmonella serogenotyping assay was used to identify a field panel of 105 Salmonella isolates. All were identified as Salmonella and 93 of 105 isolates (88.6%) were typed in full concordance with conventional serotyping. This microarray based assay is a powerful tool for serogenotyping.
format Online
Article
Text
id pubmed-3464306
institution National Center for Biotechnology Information
language English
publishDate 2012
publisher Public Library of Science
record_format MEDLINE/PubMed
spelling pubmed-34643062012-10-10 Fast DNA Serotyping and Antimicrobial Resistance Gene Determination of Salmonella enterica with an Oligonucleotide Microarray-Based Assay Braun, Sascha D. Ziegler, Albrecht Methner, Ulrich Slickers, Peter Keiling, Silke Monecke, Stefan Ehricht, Ralf PLoS One Research Article Salmonellosis caused by Salmonella (S.) belongs to the most prevalent food-borne zoonotic diseases throughout the world. Therefore, serotype identification for all culture-confirmed cases of Salmonella infection is important for epidemiological purposes. As a standard, the traditional culture method (ISO 6579:2002) is used to identify Salmonella. Classical serotyping takes 4–5 days to be completed, it is labor-intensive, expensive and more than 250 non-standardized sera are necessary to characterize more than 2,500 Salmonella serovars currently known. These technical difficulties could be overcome with modern molecular methods. We developed a microarray based serogenotyping assay for the most prevalent Salmonella serovars in Europe and North America. The current assay version could theoretically discriminate 28 O-antigens and 86 H-antigens. Additionally, we included 77 targets analyzing antimicrobial resistance genes. The Salmonella assay was evaluated with a set of 168 reference strains representing 132 serovars previously serotyped by conventional agglutination through various reference centers. 117 of 132 (81%) tested serovars showed an unique microarray pattern. 15 of 132 serovars generated a pattern which was shared by multiple serovars (e.g., S. ser. Enteritidis and S. ser. Nitra). These shared patterns mainly resulted from the high similarity of the genotypes of serogroup A and D1. Using patterns of the known reference strains, a database was build which represents the basis of a new PatternMatch software that can serotype unknown Salmonella isolates automatically. After assay verification, the Salmonella serogenotyping assay was used to identify a field panel of 105 Salmonella isolates. All were identified as Salmonella and 93 of 105 isolates (88.6%) were typed in full concordance with conventional serotyping. This microarray based assay is a powerful tool for serogenotyping. Public Library of Science 2012-10-04 /pmc/articles/PMC3464306/ /pubmed/23056321 http://dx.doi.org/10.1371/journal.pone.0046489 Text en © 2012 Braun et al http://creativecommons.org/licenses/by/4.0/ 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 properly credited.
spellingShingle Research Article
Braun, Sascha D.
Ziegler, Albrecht
Methner, Ulrich
Slickers, Peter
Keiling, Silke
Monecke, Stefan
Ehricht, Ralf
Fast DNA Serotyping and Antimicrobial Resistance Gene Determination of Salmonella enterica with an Oligonucleotide Microarray-Based Assay
title Fast DNA Serotyping and Antimicrobial Resistance Gene Determination of Salmonella enterica with an Oligonucleotide Microarray-Based Assay
title_full Fast DNA Serotyping and Antimicrobial Resistance Gene Determination of Salmonella enterica with an Oligonucleotide Microarray-Based Assay
title_fullStr Fast DNA Serotyping and Antimicrobial Resistance Gene Determination of Salmonella enterica with an Oligonucleotide Microarray-Based Assay
title_full_unstemmed Fast DNA Serotyping and Antimicrobial Resistance Gene Determination of Salmonella enterica with an Oligonucleotide Microarray-Based Assay
title_short Fast DNA Serotyping and Antimicrobial Resistance Gene Determination of Salmonella enterica with an Oligonucleotide Microarray-Based Assay
title_sort fast dna serotyping and antimicrobial resistance gene determination of salmonella enterica with an oligonucleotide microarray-based assay
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3464306/
https://www.ncbi.nlm.nih.gov/pubmed/23056321
http://dx.doi.org/10.1371/journal.pone.0046489
work_keys_str_mv AT braunsaschad fastdnaserotypingandantimicrobialresistancegenedeterminationofsalmonellaentericawithanoligonucleotidemicroarraybasedassay
AT ziegleralbrecht fastdnaserotypingandantimicrobialresistancegenedeterminationofsalmonellaentericawithanoligonucleotidemicroarraybasedassay
AT methnerulrich fastdnaserotypingandantimicrobialresistancegenedeterminationofsalmonellaentericawithanoligonucleotidemicroarraybasedassay
AT slickerspeter fastdnaserotypingandantimicrobialresistancegenedeterminationofsalmonellaentericawithanoligonucleotidemicroarraybasedassay
AT keilingsilke fastdnaserotypingandantimicrobialresistancegenedeterminationofsalmonellaentericawithanoligonucleotidemicroarraybasedassay
AT moneckestefan fastdnaserotypingandantimicrobialresistancegenedeterminationofsalmonellaentericawithanoligonucleotidemicroarraybasedassay
AT ehrichtralf fastdnaserotypingandantimicrobialresistancegenedeterminationofsalmonellaentericawithanoligonucleotidemicroarraybasedassay