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Simultaneous species identification and detection of rifampicin resistance in staphylococci by sequencing of the rpoB gene
Örebro University, School of Health and Medical Sciences.
Örebro University, School of Health and Medical Sciences.ORCID iD: 0000-0001-5939-2932
Örebro University, School of Health and Medical Sciences.
2008 (English)In: European Journal of Clinical Microbiology and Infectious Diseases, ISSN 0934-9723, E-ISSN 1435-4373, Vol. 28, no 2, p. 183-190Article in journal (Refereed) Published
Abstract [en]

In recent years, coagulase-negative staphylococci (CoNS) have been increasingly recognised as causative agents of various infections, especially in immunocompromised patients and related to implanted foreign body materials. In this study, rpoB sequencing was used for simultaneous species identification and detection of rifampicin resistance in clinical staphylococci isolates. Forty-nine (96%) out of 51 isolates, representing 17 different Staphylococcus species according to the initial phenotypic species identification, were identified to the species level using rpoB sequencing. Furthermore, the two remaining isolates were Kocuria sp. and Corynebacterium sp. respectively, according to 16S rRNA sequencing. Comparison with the phenotypic diagnostics also revealed that 8 (16%) of the 49 isolates differed regarding identified species. Discrepant analysis confirmed the result of the rpoB sequencing for all except 2 of these isolates, which could not be distinguished as single species using 16S rRNA sequencing. Regarding detection of rifampicin resistance, isolates obtained pre- and post-treatment with rifampicin were examined. These isolates comprised S. aureus (7 patients) and S. lugdunensis (1 patient). Rifampicin resistance was mainly detected following short-term treatment with rifampicin in combination with isoxazolyl-penicillin, or long-term treatment with rifampicin and ciprofloxacin. Each rifampicin-resistant isolate displayed an identical rpoB sequence as their corresponding rifampicin-susceptible isolates except for one (n = 6) or two (n = 1) nonsynonymous single nucleotide polymorphisms, or insertion of one codon (n = 1). In conclusion, rpoB sequencing is a rapid, objective and accurate method of species identification and simultaneous detection of rifampicin resistance in staphylococci.

Place, publisher, year, edition, pages
Berlin: Springer , 2008. Vol. 28, no 2, p. 183-190
National Category
Medical and Health Sciences Microbiology in the medical area
Research subject
Biomedicine
Identifiers
URN: urn:nbn:se:oru:diva-3467DOI: 10.1007/s10096-008-0604-5PubMedID: 18716806OAI: oai:DiVA.org:oru-3467DiVA, id: diva2:137764
Available from: 2008-12-08 Created: 2008-12-08 Last updated: 2018-01-13Bibliographically approved
In thesis
1. Genotypic and phenotypic characterisation of Staphylococcus epidermidis isolated from prosthetic joint infections
Open this publication in new window or tab >>Genotypic and phenotypic characterisation of Staphylococcus epidermidis isolated from prosthetic joint infections
2011 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Staphylococcus epidermidis has emerged in recent years as an important nosocomial pathogen, especially in infections associated with implanted foreign body materials (e.g., prosthetic joints and heart valves) and in individuals with a compromised immune system (e.g., cancer patients and neonates). Although rare, implant infections are long lasting and cause severe suffering for the patient that includes pain and disability and even increased mortality. One aim of the present thesis was to develop and evaluate a genetic method for species identification and simultaneous detection of rifampicin resistance in staphylococci. A second aim was to examine S. epidermidis isolated from prosthetic joint infections (PJIs) and from wrists and nares of healthy individuals regarding their antibiotic susceptibility, biofilm production, virulence factors, and epidemiology. Comparison with phenotypic diagnostics revealed that 8 (16%) of 49 isolates differed in their species identification in favour of the genetic method. In addition, mutations associated with rifampicin resistance, including two not previously reported, were possible to detect in all isolates resistant to rifampicin. Antibiotic susceptibility testing of 61 PJI isolates showed multi-drug resistance in 91%. Furthermore, the results of the synergy testing revealed that no antibiotic combination was significantly better than the others. Hence, the effects that were possible to detect were isolate dependent. To find a method for discriminating between invasive (n=61) and commensal (n=24) isolates of S. epidermidis genotypic and phenotypic characterisations of biofilm production (including the ica and aap genes), antibiotic susceptibility, virulence-related genes (such as agr and ACME) and epidemiology were performed (using multilocus sequence typing [MLST], typing of the staphylococcal chromosome cassette mec [SCCmec] and PhenePlate). Significant differences were found in antibiotic susceptibility, i.e. there was more resistance among invasive isolates. MLST sequence types (ST) ST2 and ST215 dominated the invasive isolates.

Place, publisher, year, edition, pages
Örebro: Örebro universitet, 2011. p. 117
Series
Örebro Studies in Medicine, ISSN 1652-4063 ; 53
Keywords
Staphylococcus epidermidis, prosthetic joint infections, antibiotic susceptibility, virulence factors, epidemiology, MLST, agr, SCCmec
National Category
Medical and Health Sciences
Research subject
Biomedicine
Identifiers
urn:nbn:se:oru:diva-15171 (URN)978-91-7668-793-2 (ISBN)
Public defence
2011-05-13, Wilandersalen, Universitetssjukhuset, Örebro, 09:00
Opponent
Supervisors
Available from: 2011-04-01 Created: 2011-04-01 Last updated: 2017-10-17Bibliographically approved

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Publisher's full textPubMedhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Retrieve&list_uids=18716806&dopt=Citation

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Hellmark, BengtSöderquist, BoUnemo, Magnus

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