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Rapid high-resolution melting genotyping scheme for Escherichia coli based on MLST derived single nucleotide polymorphisms

Routinely used typing methods including MLST, rep-PCR and whole genome sequencing (WGS) are time-consuming, costly, and often low throughput. Here, we describe a novel mini-MLST scheme for Eschericha coli as an alternative method for rapid genotyping. Using the proposed mini-MLST scheme, 10,946 exis...

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Autores principales: Bezdicek, Matej, Nykrynova, Marketa, Sedlar, Karel, Kralova, Stanislava, Hanslikova, Jana, Komprdova, Aja, Skutkova, Helena, Kocmanova, Iva, Mayer, Jiri, Lengerova, Martina
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8368041/
https://www.ncbi.nlm.nih.gov/pubmed/34400722
http://dx.doi.org/10.1038/s41598-021-96148-3
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author Bezdicek, Matej
Nykrynova, Marketa
Sedlar, Karel
Kralova, Stanislava
Hanslikova, Jana
Komprdova, Aja
Skutkova, Helena
Kocmanova, Iva
Mayer, Jiri
Lengerova, Martina
author_facet Bezdicek, Matej
Nykrynova, Marketa
Sedlar, Karel
Kralova, Stanislava
Hanslikova, Jana
Komprdova, Aja
Skutkova, Helena
Kocmanova, Iva
Mayer, Jiri
Lengerova, Martina
author_sort Bezdicek, Matej
collection PubMed
description Routinely used typing methods including MLST, rep-PCR and whole genome sequencing (WGS) are time-consuming, costly, and often low throughput. Here, we describe a novel mini-MLST scheme for Eschericha coli as an alternative method for rapid genotyping. Using the proposed mini-MLST scheme, 10,946 existing STs were converted into 1,038 Melting Types (MelTs). To validate the new mini-MLST scheme, in silico analysis was performed on 73,704 strains retrieved from EnteroBase resulting in discriminatory power D = 0.9465 (CI 95% 0.9726–0.9736) for mini-MLST and D = 0.9731 (CI 95% 0.9726–0.9736) for MLST. Moreover, validation on clinical isolates was conducted with a significant concordance between MLST, rep-PCR and WGS. To conclude, the great portability, efficient processing, cost-effectiveness, and high throughput of mini-MLST represents immense benefits, even when accompanied with a slightly lower discriminatory power than other typing methods. This study proved mini-MLST is an ideal method to screen and subgroup large sets of isolates and/or quick strain typing during outbreaks. In addition, our results clearly showed its suitability for prospective surveillance monitoring of emergent and high-risk E. coli clones’.
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spelling pubmed-83680412021-08-17 Rapid high-resolution melting genotyping scheme for Escherichia coli based on MLST derived single nucleotide polymorphisms Bezdicek, Matej Nykrynova, Marketa Sedlar, Karel Kralova, Stanislava Hanslikova, Jana Komprdova, Aja Skutkova, Helena Kocmanova, Iva Mayer, Jiri Lengerova, Martina Sci Rep Article Routinely used typing methods including MLST, rep-PCR and whole genome sequencing (WGS) are time-consuming, costly, and often low throughput. Here, we describe a novel mini-MLST scheme for Eschericha coli as an alternative method for rapid genotyping. Using the proposed mini-MLST scheme, 10,946 existing STs were converted into 1,038 Melting Types (MelTs). To validate the new mini-MLST scheme, in silico analysis was performed on 73,704 strains retrieved from EnteroBase resulting in discriminatory power D = 0.9465 (CI 95% 0.9726–0.9736) for mini-MLST and D = 0.9731 (CI 95% 0.9726–0.9736) for MLST. Moreover, validation on clinical isolates was conducted with a significant concordance between MLST, rep-PCR and WGS. To conclude, the great portability, efficient processing, cost-effectiveness, and high throughput of mini-MLST represents immense benefits, even when accompanied with a slightly lower discriminatory power than other typing methods. This study proved mini-MLST is an ideal method to screen and subgroup large sets of isolates and/or quick strain typing during outbreaks. In addition, our results clearly showed its suitability for prospective surveillance monitoring of emergent and high-risk E. coli clones’. Nature Publishing Group UK 2021-08-16 /pmc/articles/PMC8368041/ /pubmed/34400722 http://dx.doi.org/10.1038/s41598-021-96148-3 Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Bezdicek, Matej
Nykrynova, Marketa
Sedlar, Karel
Kralova, Stanislava
Hanslikova, Jana
Komprdova, Aja
Skutkova, Helena
Kocmanova, Iva
Mayer, Jiri
Lengerova, Martina
Rapid high-resolution melting genotyping scheme for Escherichia coli based on MLST derived single nucleotide polymorphisms
title Rapid high-resolution melting genotyping scheme for Escherichia coli based on MLST derived single nucleotide polymorphisms
title_full Rapid high-resolution melting genotyping scheme for Escherichia coli based on MLST derived single nucleotide polymorphisms
title_fullStr Rapid high-resolution melting genotyping scheme for Escherichia coli based on MLST derived single nucleotide polymorphisms
title_full_unstemmed Rapid high-resolution melting genotyping scheme for Escherichia coli based on MLST derived single nucleotide polymorphisms
title_short Rapid high-resolution melting genotyping scheme for Escherichia coli based on MLST derived single nucleotide polymorphisms
title_sort rapid high-resolution melting genotyping scheme for escherichia coli based on mlst derived single nucleotide polymorphisms
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8368041/
https://www.ncbi.nlm.nih.gov/pubmed/34400722
http://dx.doi.org/10.1038/s41598-021-96148-3
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