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      ECTyper: in silico Escherichia coli serotype and species prediction from raw and assembled whole-genome sequence data

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          Abstract

          Escherichia coli is a priority foodborne pathogen of public health concern and phenotypic serotyping provides critical information for surveillance and outbreak detection activities. Public health and food safety laboratories are increasingly adopting whole-genome sequencing (WGS) for characterizing pathogens, but it is imperative to maintain serotype designations in order to minimize disruptions to existing public health workflows. Multiple in silico tools have been developed for predicting serotypes from WGS data, including SRST2, SerotypeFinder and EToKi EBEis, but these tools were not designed with the specific requirements of diagnostic laboratories, which include: speciation, input data flexibility (fasta/fastq), quality control information and easily interpretable results. To address these specific requirements, we developed ECTyper ( https://github.com/phac-nml/ecoli_serotyping) for performing both speciation within Escherichia and Shigella , and in silico serotype prediction. We compared the serotype prediction performance of each tool on a newly sequenced panel of 185 isolates with confirmed phenotypic serotype information. We found that all tools were highly concordant, with 92–97 % for O-antigens and 98–100 % for H-antigens, and ECTyper having the highest rate of concordance. We extended the benchmarking to a large panel of 6954 publicly available E. coli genomes to assess the performance of the tools on a more diverse dataset. On the public data, there was a considerable drop in concordance, with 75–91 % for O-antigens and 62–90 % for H-antigens, and ECTyper and SerotypeFinder being the most concordant. This study highlights that in silico predictions show high concordance with phenotypic serotyping results, but there are notable differences in tool performance. ECTyper provides highly accurate and sensitive in silico serotype predictions, in addition to speciation, and is designed to be easily incorporated into bioinformatic workflows.

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          Most cited references48

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          The Sequence Alignment/Map format and SAMtools

          Summary: The Sequence Alignment/Map (SAM) format is a generic alignment format for storing read alignments against reference sequences, supporting short and long reads (up to 128 Mbp) produced by different sequencing platforms. It is flexible in style, compact in size, efficient in random access and is the format in which alignments from the 1000 Genomes Project are released. SAMtools implements various utilities for post-processing alignments in the SAM format, such as indexing, variant caller and alignment viewer, and thus provides universal tools for processing read alignments. Availability: http://samtools.sourceforge.net Contact: rd@sanger.ac.uk
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            Fast gapped-read alignment with Bowtie 2.

            As the rate of sequencing increases, greater throughput is demanded from read aligners. The full-text minute index is often used to make alignment very fast and memory-efficient, but the approach is ill-suited to finding longer, gapped alignments. Bowtie 2 combines the strengths of the full-text minute index with the flexibility and speed of hardware-accelerated dynamic programming algorithms to achieve a combination of high speed, sensitivity and accuracy.
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              Basic local alignment search tool.

              A new approach to rapid sequence comparison, basic local alignment search tool (BLAST), directly approximates alignments that optimize a measure of local similarity, the maximal segment pair (MSP) score. Recent mathematical results on the stochastic properties of MSP scores allow an analysis of the performance of this method as well as the statistical significance of alignments it generates. The basic algorithm is simple and robust; it can be implemented in a number of ways and applied in a variety of contexts including straightforward DNA and protein sequence database searches, motif searches, gene identification searches, and in the analysis of multiple regions of similarity in long DNA sequences. In addition to its flexibility and tractability to mathematical analysis, BLAST is an order of magnitude faster than existing sequence comparison tools of comparable sensitivity.
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                Author and article information

                Journal
                Microb Genom
                Microb Genom
                mgen
                mgen
                Microbial Genomics
                Microbiology Society
                2057-5858
                2021
                3 December 2021
                3 December 2021
                : 7
                : 12
                : 000728
                Affiliations
                [ 1] departmentNational Microbiology Laboratory , Public Health Agency of Canada , Guelph, ON, Canada
                [ 2] departmentNational Centre for Animal Diseases , Canadian Food Inspection Agency , Lethbridge, Canada
                [ 3] departmentNational Microbiology Laboratory , Public Health Agency of Canada , Lethbridge, AB, Canada
                [ 4] departmentNational Microbiology Laboratory , Public Health Agency of Canada , Toronto, ON, Canada
                [ 5] departmentNational Microbiology Laboratory , Public Health Agency of Canada , Winnipeg, MB, Canada
                Author notes
                *Correspondence: Kyrylo Bessonov, kyrylo.bessonov@ 123456canada.ca
                [†]

                These authors contributed equally to this work

                [‡]

                These authors also contributed equally to this work

                Author information
                https://orcid.org/0000-0002-9579-3183
                https://orcid.org/0000-0003-1162-257X
                https://orcid.org/0000-0002-8534-5694
                https://orcid.org/0000-0002-5133-3409
                https://orcid.org/0000-0001-6064-405X
                https://orcid.org/0000-0001-5889-7429
                https://orcid.org/0000-0002-1860-8861
                https://orcid.org/0000-0003-2925-8710
                Article
                000728
                10.1099/mgen.0.000728
                8767331
                34860150
                c4b8b806-7356-4e57-ab20-e986e470faa6
                © 2021 Crown Copyright

                This is an open-access article distributed under the terms of the Creative Commons Attribution License.

                History
                : 29 April 2021
                : 25 October 2021
                Funding
                Funded by: Public Health Agency of Canada
                Award Recipient : KyryloBessonov
                Categories
                Research Articles
                Pathogens and Epidemiology
                Custom metadata
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                enteric pathogens,e. coli, in silico serotyping,public health,serotyping

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