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      Core Genome Multilocus Sequence Typing Scheme for Stable, Comparative Analyses of Campylobacter jejuni and C. coli Human Disease Isolates

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          ABSTRACT

          Human campylobacteriosis, caused by Campylobacter jejuni and C. coli, remains a leading cause of bacterial gastroenteritis in many countries, but the epidemiology of campylobacteriosis outbreaks remains poorly defined, largely due to limitations in the resolution and comparability of isolate characterization methods. Whole-genome sequencing (WGS) data enable the improvement of sequence-based typing approaches, such as multilocus sequence typing (MLST), by substantially increasing the number of loci examined. A core genome MLST (cgMLST) scheme defines a comprehensive set of those loci present in most members of a bacterial group, balancing very high resolution with comparability across the diversity of the group. Here we propose a set of 1,343 loci as a human campylobacteriosis cgMLST scheme (v1.0), the allelic profiles of which can be assigned to core genome sequence types. The 1,343 loci chosen were a subset of the 1,643 loci identified in the reannotation of the genome sequence of C. jejuni isolate NCTC 11168, chosen as being present in >95% of draft genomes of 2,472 representative United Kingdom campylobacteriosis isolates, comprising 2,207 (89.3%) C. jejuni isolates and 265 (10.7%) C. coli isolates. Validation of the cgMLST scheme was undertaken with 1,478 further high-quality draft genomes, containing 150 or fewer contiguous sequences, from disease isolate collections: 99.5% of these isolates contained ≥95% of the 1,343 cgMLST loci. In addition to the rapid and effective high-resolution analysis of large numbers of diverse isolates, the cgMLST scheme enabled the efficient identification of very closely related isolates from a well-defined single-source campylobacteriosis outbreak.

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          The genome sequence of the food-borne pathogen Campylobacter jejuni reveals hypervariable sequences.

          Campylobacter jejuni, from the delta-epsilon group of proteobacteria, is a microaerophilic, Gram-negative, flagellate, spiral bacterium-properties it shares with the related gastric pathogen Helicobacter pylori. It is the leading cause of bacterial food-borne diarrhoeal disease throughout the world. In addition, infection with C. jejuni is the most frequent antecedent to a form of neuromuscular paralysis known as Guillain-Barré syndrome. Here we report the genome sequence of C. jejuni NCTC11168. C. jejuni has a circular chromosome of 1,641,481 base pairs (30.6% G+C) which is predicted to encode 1,654 proteins and 54 stable RNA species. The genome is unusual in that there are virtually no insertion sequences or phage-associated sequences and very few repeat sequences. One of the most striking findings in the genome was the presence of hypervariable sequences. These short homopolymeric runs of nucleotides were commonly found in genes encoding the biosynthesis or modification of surface structures, or in closely linked genes of unknown function. The apparently high rate of variation of these homopolymeric tracts may be important in the survival strategy of C. jejuni.
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            MLST revisited: the gene-by-gene approach to bacterial genomics.

            Multilocus sequence typing (MLST) was proposed in 1998 as a portable sequence-based method for identifying clonal relationships among bacteria. Today, in the whole-genome era of microbiology, the need for systematic, standardized descriptions of bacterial genotypic variation remains a priority. Here, to meet this need, we draw on the successes of MLST and 16S rRNA gene sequencing to propose a hierarchical gene-by-gene approach that reflects functional and evolutionary relationships and catalogues bacteria 'from domain to strain'. Our gene-based typing approach using online platforms such as the Bacterial Isolate Genome Sequence Database (BIGSdb) allows the scalable organization and analysis of whole-genome sequence data.
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              Transforming clinical microbiology with bacterial genome sequencing.

              Whole-genome sequencing of bacteria has recently emerged as a cost-effective and convenient approach for addressing many microbiological questions. Here, we review the current status of clinical microbiology and how it has already begun to be transformed by using next-generation sequencing. We focus on three essential tasks: identifying the species of an isolate, testing its properties, such as resistance to antibiotics and virulence, and monitoring the emergence and spread of bacterial pathogens. We predict that the application of next-generation sequencing will soon be sufficiently fast, accurate and cheap to be used in routine clinical microbiology practice, where it could replace many complex current techniques with a single, more efficient workflow.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                J Clin Microbiol
                J. Clin. Microbiol
                jcm
                jcm
                JCM
                Journal of Clinical Microbiology
                American Society for Microbiology (1752 N St., N.W., Washington, DC )
                0095-1137
                1098-660X
                26 April 2017
                23 June 2017
                July 2017
                23 June 2017
                : 55
                : 7
                : 2086-2097
                Affiliations
                [a ]Department of Zoology, University of Oxford, Oxford, United Kingdom
                [b ]NIHR Health Protection Research Unit in Gastrointestinal Infections, University of Oxford, Oxford, United Kingdom
                [c ]Warwick Medical School, University of Warwick, Coventry, United Kingdom
                University of Iowa College of Medicine
                Author notes
                Address correspondence to Alison J. Cody, alison.cody@ 123456zoo.ox.ac.uk .

                Citation Cody AJ, Bray JE, Jolley KA, McCarthy ND, Maiden MCJ. 2017. Core genome multilocus sequence typing scheme for stable, comparative analyses of Campylobacter jejuni and C. coli human disease isolates. J Clin Microbiol 55:2086–2097. https://doi.org/10.1128/JCM.00080-17.

                Author information
                http://orcid.org/0000-0001-6321-5138
                Article
                00080-17
                10.1128/JCM.00080-17
                5483910
                28446571
                6529b398-4d81-4028-9b41-1770e7f83ab6
                Copyright © 2017 Cody et al.

                This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.

                History
                : 13 January 2017
                : 17 February 2017
                : 13 April 2017
                Page count
                supplementary-material: 3, Figures: 5, Tables: 2, Equations: 0, References: 39, Pages: 12, Words: 6908
                Funding
                Funded by: Department for Environment, Food and Rural Affairs (Defra) https://doi.org/10.13039/501100000277
                Award ID: OZ0624
                Award Recipient : Alison J. Cody
                Funded by: DH | National Institute for Health Research (NIHR) https://doi.org/10.13039/501100000272
                Award ID: 10038
                Award Recipient : Alison J. Cody Award Recipient : Noel D. McCarthy Award Recipient : Martin C.J. Maiden
                Funded by: Food Standards Agency (FSA) https://doi.org/10.13039/501100000354
                Award ID: OZ0624
                Award Recipient : Alison J. Cody
                Funded by: Wellcome https://doi.org/10.13039/100010269
                Award ID: 104992
                Award Recipient : James E. Bray Award Recipient : Keith A. Jolley
                Funded by: Wellcome https://doi.org/10.13039/100010269
                Award ID: 087622
                Award Recipient : Martin C.J. Maiden
                Funded by: Wellcome https://doi.org/10.13039/100010269
                Award ID: 072782MA
                Award Recipient : Noel D. McCarthy
                Categories
                Epidemiology
                Custom metadata
                July 2017

                Microbiology & Virology
                campylobacter,molecular epidemiology,whole-genome sequencing,core genome

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