Staff Publications

Staff Publications

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    'Staff publications' is the digital repository of Wageningen University & Research

    'Staff publications' contains references to publications authored by Wageningen University staff from 1976 onward.

    Publications authored by the staff of the Research Institutes are available from 1995 onwards.

    Full text documents are added when available. The database is updated daily and currently holds about 240,000 items, of which 72,000 in open access.

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Record number 487958
Title Cell-Free Propagation of Coxiella burnetii Does Not Affect Its Relative Virulence
Author(s) Kuley, R.; Smith, H.E.; Frangoulidis, D.; Smits, M.A.; Roest, H.I.J.; Bossers, A.
Source PLoS ONE 10 (2015)3. - ISSN 1932-6203 - 16 p.
DOI https://doi.org/10.1371/journal.pone.0121661
Department(s) Host-Microbe Interactomics
Infection Biology
Wageningen Livestock Research
Animal Breeding & Genomics
Bacteriology & Epidemiology
Publication type Refereed Article in a scientific journal
Publication year 2015
Keyword(s) real-time pcr - q-fever - ethidium monoazide - phase-ii - propidium monoazide - dead cells - t-cells - lipopolysaccharide - infection - strains
Abstract Q fever is caused by the obligate intracellular bacterium Coxiella burnetii. In vitro growth of the bacterium is usually limited to viable eukaryotic host cells imposing experimental constraints for molecular studies, such as the identification and characterisation of major virulence factors. Studies of pathogenicity may benefit from the recent development of an extracellular growth medium for C. burnetii. However, it is crucial to investigate the consistency of the virulence phenotype of strains propagated by the two fundamentally different culturing systems. In the present study, we assessed the viability of C. burnetii and the lipopolysaccaride (LPS) encoding region of the bacteria in both culture systems as indirect but key parameters to the infection potential of C. burnetii. Propidium monoazide (PMA) treatment-based real-time PCR was used for enumeration of viable C. burnetii which were validated by fluorescent infectious focus forming unit counting assays. Furthermore, RNA isolated from C. burnetiipropagated in both the culture systems was examined for LPS-related gene expression. All thus far known LPS-related genes were found to be expressed in early passages in both culturing systems indicating the presence of predominantly the phase I form of C. burnetii. Finally, we used immune-competent mice to provide direct evidence, that the relative virulence of different C. burnetii strains is essentially the same for both axenic and cell-based methods of propagation.
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