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      Nosocomial infection due to Enterococcus cecorum identified by MALDI-TOF MS and Vitek 2 from a blood culture of a septic patient

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          Abstract

          We report the case of a nosocomial infection due to Enterococcus cecorum isolated from a blood culture of a 75-year-old septic male patient. Matrix-assisted laser desorption–ionization time-of-flight mass spectrometry (MALDI-TOF MS) and Vitek 2 succeeded in identification of the isolate.

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          16S ribosomal DNA amplification for phylogenetic study.

          D J R Lane, D A Pelletier, S M Barns … (1991)
          A set of oligonucleotide primers capable of initiating enzymatic amplification (polymerase chain reaction) on a phylogenetically and taxonomically wide range of bacteria is described along with methods for their use and examples. One pair of primers is capable of amplifying nearly full-length 16S ribosomal DNA (rDNA) from many bacterial genera; the additional primers are useful for various exceptional sequences. Methods for purification of amplified material, direct sequencing, cloning, sequencing, and transcription are outlined. An obligate intracellular parasite of bovine erythrocytes, Anaplasma marginale, is used as an example; its 16S rDNA was amplified, cloned, sequenced, and phylogenetically placed. Anaplasmas are related to the genera Rickettsia and Ehrlichia. In addition, 16S rDNAs from several species were readily amplified from material found in lyophilized ampoules from the American Type Culture Collection. By use of this method, the phylogenetic study of extremely fastidious or highly pathogenic bacterial species can be carried out without the need to culture them. In theory, any gene segment for which polymerase chain reaction primer design is possible can be derived from a readily obtainable lyophilized bacterial culture.
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            Characterization of universal small-subunit rRNA hybridization probes for quantitative molecular microbial ecology studies.

            Elizabeth Alm, Peter D. Stahl, L Raskin … (1996)
            Universal oligonucleotide hybridization probes targeting the small-subunit rRNA are commonly used to quantify total microbial representation in environmental samples. Universal probes also serve to normalize results obtained with probes targeting specific phylogenetic groups of microorganisms. In this study, six universal probes were evaluated for stability of probe-target duplexes by using rRNA from nine organisms representing the three domains of Bacteria, Archaea, and Eucarya. Domain-specific variations in dissociation temperatures were observed for all probes. This could lead to a significant bias when these probes are used to quantify microbial populations in environmental samples. We suggest lowering the posthybridization wash stringency for two of the universal probes (S-*-Univ-1390-a-A-18 and S-*-Univ-1392-a-A-15) examined. These two probes were evaluated with traditional and modified hybridization conditions to characterize defined mixtures of rRNAs extracted from pure cultures and rRNA samples obtained from anaerobic digester samples. Probe S-*-Univ-1390-a-A-18 provided excellent estimations of domain-level community composition of these samples and is recommended for future use in microbial ecology studies.
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              Enterococcus cecorum infections in broiler breeders and their offspring: molecular epidemiology.

              M J Kense, Gijs Landman (2011)
              Increased mortality and problems with lameness were reported in Dutch broiler flocks from the year 2008 onwards. Therefore, a field inventory, including 10 affected broiler flocks, nine corresponding broiler breeder flocks and five hatcheries, was carried out. The onset of clinical signs (lameness and increased mortality) started at about 2 weeks of age. The flock mortality varied from 3.1 to 8.1% at slaughter. Post-mortem lesions of broiler flocks were characterized by the occurrence of pericarditis/hydropericardium, arthritis and femoral head necrosis. Enterococcus cecorum was isolated from approximately 30% of the lesions. In the broiler breeders, E. cecorum was not isolated from any of the lesions. However, it was isolated from 31 out of 65 (47%) cloacal swabs, from two out of 65 (3%) oviduct samples, from one out 65 (1.5%) bone marrow samples and from two out of 25 (8%) blood samples. E. cecorum was not isolated from the air samples or dead-in-shell originating from the hatcheries involved. In total, 78 isolates were subjected to further typing by means of tRNA intergenic spacer PCR and confirmed as E. cecorum. The genetic relatedness of these cocci was subsequently studied using pulsed-field gel electrophoresis. The banding patterns of approximately 68% of E. cecorum isolates originating from parent stock flocks were clonal to one or more isolates of the same or other parent flocks. In contrast, isolates originating from their diseased offspring showed much greater genetic variation. Therefore, the vertical transmission of E. cecorum could not be demonstrated.
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                Author and article information

                Journal
                Journal ID (nlm-ta): Eur J Microbiol Immunol (Bp)
                Journal ID (iso-abbrev): Eur J Microbiol Immunol (Bp)
                Journal ID (publisher-id): EUJMI
                Title: European Journal of Microbiology & Immunology
                Publisher: Akadémiai Kiadó (Budapest )
                ISSN (Print): 2062-509X
                ISSN (Electronic): 2062-8633
                Publication date (Electronic): 18 June 2015
                Publication date Collection: June 2015
                Volume: 5
                Issue: 2
                Pages: 177-179
                Affiliations
                [1 ]Institute of Medical Microbiology, Virology, and Hygiene, Rostock University Hospital , Rostock, Germany
                [2 ] Department of Pneumology and Critical Care Medicine, Rostock University Hospital , Rostock, Germany
                Author notes
                * Institute of Medical Microbiology, Virology and Hygiene, Rostock University Hospital, Schillingallee 70, 18057 Rostock, Germany; philipp.warnke@ 123456med.uni-rostock.de
                Article
                DOI: 10.1556/1886.2015.00008
                PMC ID: 4500070
                PubMed ID: 26185687
                SO-VID: 8e1783a8-bffe-492c-ab8c-cfda1b84cb03
                Copyright © © 2015, The Author(s)
                License:

                This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License ( http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                Date received : 25 February 2015
                Date accepted : 02 March 2015
                Page count
                Figures: 0, Tables: 1, Equations: 0, References: 12, Pages: 3
                Categories
                Subject: Case Study

                Keywords: enterococcus cecorum,nosocomial infection,maldi-tof,mass spectrometry,vitek 2
                Data availability:
                Keywords: enterococcus cecorum, nosocomial infection, maldi-tof, mass spectrometry, vitek 2

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