Cell-free propagation of Coxiella burnetii does not affect its relative virulence

PloS One
Runa KuleyAlex Bossers

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 predomi...Continue Reading

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Citations

Jul 16, 2016·Future Microbiology·Charles L LarsonMatteo Bonazzi
Jan 6, 2017·Vector Borne and Zoonotic Diseases·Marcella MoriAna Sofia Santos
Nov 24, 2017·International Journal of Molecular Sciences·Prasad AbnaveEric Ghigo
Jun 22, 2018·Current Protocols in Microbiology·Savannah E SanchezAnders Omsland
Jun 18, 2017·Frontiers in Cellular and Infection Microbiology·Eduardo Vallejo EsquerraAnders Omsland

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Datasets Mentioned

BETA
AY502846
AE016828.2
CP000733.1
CP001020.1
CP001019.1
CP001021.1
RSA493

Methods Mentioned

BETA
genetic modification
PCR
RSA
PMA
PCRs
feature extraction
PMA-PCR

Software Mentioned

Agilent
ImageQuant
Bowtie2
GeneSpring

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Antifungals (ASM)

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