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      Molecular characterisation of methicillin-resistant Staphylococcus aureus from chronically colonised cystic fibrosis paediatric patients in Brazil

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      1 , 1 , 2 , 1 , 1
      Epidemiology and Infection
      Cambridge University Press
      MRSA, Cystic fibrosis, PFGE, SCCmec, spa typing, Staphylococcus aureus

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          Abstract

          Persistent methicillin-resistant Staphylococcus aureus (MRSA) infection in cystic fibrosis (CF) patients has been associated with a more rapid decline in lung function, increased hospitalisation and mortality. The aim of this study was to evaluate the clonal relationships among 116 MRSA isolates from 12 chronically colonised CF pediatric patients over a 6-year period in a Rio de Janeiro CF specialist centre. Isolates were characterised by antimicrobial resistance, SCC mec type, presence of Panton-Valentine Leukocidin (PVL) genes and grouped according to DNA macrorestriction profile by pulsed-field gel electrophoresis (PFGE) and spa gene type. High resistance rates were detected for erythromycin (78%) and ciprofloxacin (50%) and SCC mec IV was the most common type (72.4%). Only 8.6% of isolates were PVL positive. High genetic diversity was evident by PFGE (39 pulsotypes) and of nine that were identified spa types, t002 (53.1%) and t539 (14.8%) were the most prevalent. We conclude that the observed homogeneity of spa types within patients over the study period demonstrates the persistence of such strain lineages throughout the course of chronic lung infection.

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          Most cited references31

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          Evaluation of protein A gene polymorphic region DNA sequencing for typing of Staphylococcus aureus strains.

          Three hundred and twenty isolates of Staphylococcus aureus were typed by DNA sequence analysis of the X region of the protein A gene (spa). spa typing was compared to both phenotypic and molecular techniques for the ability to differentiate and categorize S. aureus strains into groups that correlate with epidemiological information. Two previously characterized study populations were examined. A collection of 59 isolates (F. C. Tenover, R. Arbeit, G. Archer, J. Biddle, S. Byrne, R. Goering, G. Hancock, G. A. Hébert, B. Hill, R. Hollis, W. R. Jarvis, B. Kreiswirth, W. Eisner, J. Maslow, L. K. McDougal, J. M. Miller, M. Mulligan, and M. A. Pfaller, J. Clin. Microbiol. 32:407-415, 1994) from the Centers for Disease Control and Prevention (CDC) was used to test for the ability to discriminate outbreak from epidemiologically unrelated strains. A separate collection of 261 isolates form a multicenter study (R. B. Roberts, A. de Lencastre, W. Eisner, E. P. Severina, B. Shopsin, B. N. Kreiswirth, and A. Tomasz, J. Infect. Dis. 178:164-171, 1998) of methicillin-resistant S. aureus in New York City (NYC) was used to compare the ability of spa typing to group strains along clonal lines to that of the combination of pulsed-field gel electrophoresis and Southern hybridization. In the 320 isolates studied, spa typing identified 24 distinct repeat types and 33 different strain types. spa typing distinguished 27 of 29 related strains and did not provide a unique fingerprint for 4 unrelated strains from the four outbreaks of the CDC collection. In the NYC collection, spa typing provided a clonal assignment for 185 of 195 strains within the five major groups previously described. spa sequencing appears to be a highly effective rapid typing tool for S. aureus that, despite some expense of specificity, has significant advantages in terms of speed, ease of use, ease of interpretation, and standardization among laboratories.
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            SCC mec Finder, a Web-Based Tool for Typing of Staphylococcal Cassette Chromosome mec in Staphylococcus aureus Using Whole-Genome Sequence Data

            SCCmec in MRSA is acknowledged to be of importance not only because it contains the mecA or mecC gene but also for staphylococcal adaptation to different environments, e.g., in hospitals, the community, and livestock. Typing of SCCmec by PCR techniques has, because of its heterogeneity, been challenging, and whole-genome sequencing has only partially solved this since no good bioinformatic tools have been available. In this article, we describe the development of a new bioinformatic tool, SCCmecFinder, that includes most of the needs for infection control professionals and researchers regarding the interpretation of SCCmec elements. The software detects all of the SCCmec elements accepted by the International Working Group on the Classification of Staphylococcal Cassette Chromosome Elements, and users will be prompted if diverging and potential new elements are uploaded. Furthermore, SCCmecFinder will be curated and updated as new elements are found and it is easy to use and freely accessible.
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              A new multiplex PCR for easy screening of methicillin-resistant Staphylococcus aureus SCCmec types I-V.

              A multiplex PCR with four primer-pairs was designed to identify the five main known SCCmec types. A clear and easily discriminated band pattern was obtained for all five types. The SCCmec type was identified for 98% of 312 clinical isolates of methicillin-resistant Staphylococcus aureus (MRSA). SCCmec type IV was by far the most common SCCmec type among both hospital- and community-acquired MRSA isolates in Denmark.
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                Author and article information

                Journal
                Epidemiol Infect
                Epidemiol. Infect
                HYG
                Epidemiology and Infection
                Cambridge University Press (Cambridge, UK )
                0950-2688
                1469-4409
                2020
                26 May 2020
                : 148
                : e149
                Affiliations
                [1 ]Departamento de Microbiologia, Imunologia e Parasitologia, Faculdade de Ciências Médicas, Universidade do Estado do Rio de Janeiro , Boulevard 28 de Setembro, 87, fundos, 3° andar – Vila Isabel, Rio de Janeiro, 20551-030, Brazil
                [2 ]Departamento de Pneumologia Pediátrica, Instituto Nacional da Saúde da Mulher da Criança e do Adolescente Fernandes Figueira, Fundação Oswaldo Cruz , Av. Rui Barbosa, 716 – Flamengo, Rio de Janeiro 20021-140, Brazil
                Author notes
                Author for correspondence: R. S. Leão, E-mail: robson.leao@ 123456uerj.br
                Author information
                https://orcid.org/0000-0003-3341-101X
                https://orcid.org/0000-0003-0636-1520
                Article
                S0950268820001156
                10.1017/S0950268820001156
                7374800
                32450935
                95d5b180-9759-489f-a79b-50d540759963
                © The Author(s) 2020

                This is an Open Access article, distributed under the terms of the Creative Commons Attribution licence ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted re-use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 13 January 2020
                : 16 May 2020
                : 18 May 2020
                Page count
                Figures: 1, Tables: 2, References: 35, Pages: 7
                Categories
                Original Paper

                Public health
                mrsa,cystic fibrosis,pfge,sccmec,spa typing,staphylococcus aureus
                Public health
                mrsa, cystic fibrosis, pfge, sccmec, spa typing, staphylococcus aureus

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