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      An Expanded Multilocus Sequence Typing Scheme for Propionibacterium acnes: Investigation of ‘Pathogenic’, ‘Commensal’ and Antibiotic Resistant Strains

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          Abstract

          The Gram-positive bacterium Propionibacterium acnes is a member of the normal human skin microbiota and is associated with various infections and clinical conditions. There is tentative evidence to suggest that certain lineages may be associated with disease and others with health. We recently described a multilocus sequence typing scheme (MLST) for P. acnes based on seven housekeeping genes ( http://pubmlst.org/pacnes). We now describe an expanded eight gene version based on six housekeeping genes and two ‘putative virulence’ genes (eMLST) that provides improved high resolution typing (91eSTs from 285 isolates), and generates phylogenies congruent with those based on whole genome analysis. When compared with the nine gene MLST scheme developed at the University of Bath, UK, and utilised by researchers at Aarhus University, Denmark, the eMLST method offers greater resolution. Using the scheme, we examined 208 isolates from disparate clinical sources, and 77 isolates from healthy skin. Acne was predominately associated with type IA 1 clonal complexes CC1, CC3 and CC4; with eST1 and eST3 lineages being highly represented. In contrast, type IA 2 strains were recovered at a rate similar to type IB and II organisms. Ophthalmic infections were predominately associated with type IA 1 and IA 2 strains, while type IB and II were more frequently recovered from soft tissue and retrieved medical devices. Strains with rRNA mutations conferring resistance to antibiotics used in acne treatment were dominated by eST3, with some evidence for intercontinental spread. In contrast, despite its high association with acne, only a small number of resistant CC1 eSTs were identified. A number of eSTs were only recovered from healthy skin, particularly eSTs representing CC72 (type II) and CC77 (type III). Collectively our data lends support to the view that pathogenic versus truly commensal lineages of P. acnes may exist. This is likely to have important therapeutic and diagnostic implications.

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          eBURST: inferring patterns of evolutionary descent among clusters of related bacterial genotypes from multilocus sequence typing data.

          The introduction of multilocus sequence typing (MLST) for the precise characterization of isolates of bacterial pathogens has had a marked impact on both routine epidemiological surveillance and microbial population biology. In both fields, a key prerequisite for exploiting this resource is the ability to discern the relatedness and patterns of evolutionary descent among isolates with similar genotypes. Traditional clustering techniques, such as dendrograms, provide a very poor representation of recent evolutionary events, as they attempt to reconstruct relationships in the absence of a realistic model of the way in which bacterial clones emerge and diversify to form clonal complexes. An increasingly popular approach, called BURST, has been used as an alternative, but present implementations are unable to cope with very large data sets and offer crude graphical outputs. Here we present a new implementation of this algorithm, eBURST, which divides an MLST data set of any size into groups of related isolates and clonal complexes, predicts the founding (ancestral) genotype of each clonal complex, and computes the bootstrap support for the assignment. The most parsimonious patterns of descent of all isolates in each clonal complex from the predicted founder(s) are then displayed. The advantages of eBURST for exploring patterns of evolutionary descent are demonstrated with a number of examples, including the simple Spain(23F)-1 clonal complex of Streptococcus pneumoniae, "population snapshots" of the entire S. pneumoniae and Staphylococcus aureus MLST databases, and the more complicated clonal complexes observed for Campylobacter jejuni and Neisseria meningitidis.
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            LIAN 3.0: detecting linkage disequilibrium in multilocus data. Linkage Analysis.

            LIAN is a program to test the null hypothesis of linkage equilibrium for multilocus data. LIAN incorporates both a Monte Carlo method as well as a novel algebraic method to carry out the hypothesis test. The program further returns the genetic diversity of the sample and the pairwise distances between its members.
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              The complete genome sequence of Propionibacterium acnes, a commensal of human skin.

              Propionibacterium acnes is a major inhabitant of adult human skin, where it resides within sebaceous follicles, usually as a harmless commensal although it has been implicated in acne vulgaris formation. The entire genome sequence of this Gram-positive bacterium encodes 2333 putative genes and revealed numerous gene products involved in degrading host molecules, including sialidases, neuraminidases, endoglycoceramidases, lipases, and pore-forming factors. Surface-associated and other immunogenic factors have been identified, which might be involved in triggering acne inflammation and other P. acnes-associated diseases.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, USA )
                1932-6203
                2012
                30 July 2012
                : 7
                : 7
                : e41480
                Affiliations
                [1 ]Centre for Infection and Immunity, School of Medicine, Dentistry and Biomedical Sciences, Queen’s University, Belfast, United Kingdom
                [2 ]Institute of Biochemistry, Biological Research Centre of the Hungarian Academy of Sciences, Szeged, Hungary
                [3 ]School of Life Sciences, University of Warwick, Coventry, United Kingdom
                [4 ]Department of Molecular and Medical Pharmacology, Crump Institute for Molecular Imaging, University of California Los Angeles, Los Angeles, California, United States of America
                [5 ]Department of Dermatology, Harrogate and District NHS Foundation Trust, Harrogate, United Kingdom
                [6 ]Faculty of Biological Sciences, University of Leeds, Leeds, United Kingdom
                [7 ]Department of Laboratory Medicine, Karolinska University Hospital Huddinge, Karolinska Institute, Stockholm, Sweden
                The University of Hong Kong, Hong Kong
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                Conceived and designed the experiments: AMD. Performed the experiments: AMD EB AG IN. Analyzed the data: AMD EB SP. Contributed reagents/materials/analysis tools: ST HL AE JC CN. Wrote the paper: AMD SP.

                Article
                PONE-D-12-12798
                10.1371/journal.pone.0041480
                3408437
                22859988
                16b6ab02-071c-45cc-a8c3-2cee27a17330
                Copyright @ 2012

                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 credited.

                History
                : 29 April 2012
                : 21 June 2012
                Page count
                Pages: 14
                Funding
                The work was supported by funding from the R & D Office of the Health and Personal Social Services Northern Ireland (grant number HSCNI 9.41; http://www.dhsspsni.gov.uk/ahp_research) and The Prostate Cancer Charity, United Kingdom (grant number 110831; http://www.prostate-cancer.org.uk/). IN: Hungarian National Office for Research and Technology Teller program OMFB-00441/2007, French-Hungarian Associated European Laboratory (LEA) SkinChroma OMFB-00272/2009, and TAMOP-4.2.1.B-10/2/KONV-2010-0002. AG: Micropathology Ltd and the Royal Commission for the Exhibition of 1851. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Biology
                Microbiology
                Bacterial Pathogens
                Gram Positive
                Bacteriology
                Bacterial Taxonomy
                Medical Microbiology
                Population Biology
                Epidemiology
                Infectious Disease Epidemiology
                Medicine
                Dermatology
                Dermatologic Pathology
                Epidemiology
                Infectious Disease Epidemiology

                Uncategorized
                Uncategorized

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